Treatment of cancer, inflammatory disease, and autoimmune disease

ABSTRACT

A method of treating cancer, inflammatory disease, and autoimmune disease by administering to a subject in need thereof an effective amount of one or more 1,5-dipenylpenta-1,4-dien-3-one compounds. The compounds feature either or both of the phenyl rings being substituted with hydroxyl, diethyl(2-alkoxyethyl)amine, 1-(2-alkoxyethyl)piperidine, sulfonate, phosphinate, or phosphate.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/210,728, filed on Aug. 16, 2011, which claims the benefit U.S.Provisional Application No. 61/375,534, filed on Aug. 20, 2010. Thecontents of both applications are hereby incorporated by reference intheir entirety.

BACKGROUND OF THE INVENTION

Signal Transducer and Activator of Transcription (STAT) proteins aretranscription factors that mediate cellular responses to growth factors.These proteins are activated via tyrosine phosphorylation by growthfactor receptor-associated tyrosine kinases. Activated STAT proteinspromote cell survival and proliferation. It is now well established thatpersistent activation of STAT3 or STAT5 promotes cell growth, invasion,and metastasis of both solid and hematopoietic cancers. See, e.g.,Expert Opin. Investig. Drugs, 2009, 18(1): 45-56.

In normal lymphoid cells, STAT proteins, e.g., STAT3, also mediatecellular responses to cytokines, such as interleukin-6 (IL-6), viacytokine receptor-associated Janus kinases (JAKs). See, e.g., Neoplasia2008, 10: 287-297. IL-6 is abnormally elevated in patients sufferingfrom hematopoietic cancers, inflammatory diseases, autoimmune diseases,and postmenopausal osteoporosis. Inhibiting cellular action of IL-6,e.g., by inducing STAT3 proteins degradation, has attributed totreatment of these diseases.

As such, compounds that deactivate STAT proteins can be used to treatvarious cancers, inflammatory diseases, and autoimmune diseases.

SUMMARY OF THE INVENTION

This invention is based on a discovery that a group of compounds havinga backbone of 1,5-diphenyl-penta-1,4-dien-3-one deactivate STATproteins.

One aspect of this invention relates to a compound of Formula (I):

wherein each of X and Y, independently, is H, alkyl, or halo, or X and Ytogether are

—CH₂—, —(CH₂)₂—, —(CH₂)CR_(a)R_(b)(CH₂)—, —(CH₂)NR_(a)(CH₂)—, or—(CH₂)O(CH₂)—, each of R_(a) and R_(b), independently, being H, alkyl,C(O)-alkyl, C(O)-cycloalkyl, C(O)—NH-alkyl, or C(O)—NH-cycloalkyl; andeach of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′),independently, is H, alkyl (e.g., alkyl substituted with halo orSO₂R_(d)), halo, OH, R_(c)—O—, R_(d)S(O)₂—O—, or (R_(d))₂P(O)—O—, R_(c)being unsubstituted alkyl or alkyl substituted with halo, OH, alkoxy,amino, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, and R_(d)being H, OH, alkyl, alkoxy, amino, or aryl; in which R¹ is differentfrom R^(1′) or R^(5′), R² is different from R^(2′) or R^(4′), R³ isdifferent from R^(3′), R⁴ is different from R^(2′) or R^(4′), or R⁵ isdifferent from R^(1′) or R^(5′).

Referring to Formula (I), a subset of the compounds have one or more ofthe following features: R² is OH or R² is R_(d)S(O)₂—O—, or(R_(d))₂P(O)—O— (R_(d) is H, OH, alkyl, alkoxy, amino, or aryl, e.g.,R_(d) is ethyl); R^(2′) is R_(c)—O— (R_(c) is alkyl substituted withamino); each of X and Y is H or X and Y together are —(CH₂)NR_(a)(CH₂)—;and R_(a) is C(O)—R, C(O)NRR′, or alkyl substituted with cycloalkyl(each of R and R′, independently, is alkyl or cycloalkyl).

Another aspect of this invention relates to a compound of Formula (II):

wherein each of X and Y, independently, is H, alkyl, or halo, or X and Ytogether are

—CH₂—, —(CH₂)₂—, —(CH₂)CR_(a)R_(b)(CH₂)—, —(CH₂)NR_(a)(CH₂)—, or—(CH₂)O(CH₂)—, each of R_(a) and R_(b), independently, being H, alkyl,C(O)-alkyl, C(O)-cycloalkyl, C(O)—NH-alkyl, or C(O)—NH-cycloalkyl; andeach of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′),independently, is H, alkyl (e.g., alkyl substituted with halo orSO₂R_(d)), halo, OH, R_(c)—O—, R_(d)S(O)₂—O—, or (R_(d))₂P(O)—O—, R_(c)being unsubstituted alkyl or alkyl substituted with halo, OH, alkoxy,amino, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, and R_(d)being H, OH, alkyl, aloxyl, amino, or aryl; in which at least one of R¹,R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′) isR_(d)S(O)₂—O—, (R_(d))₂P(O)—O—, or (R_(d)O)₂P(O)—O—.

Referring to Formula (II), a subset of the compounds have one or more ofthe following features: each of X and Y is H; R² is R_(d)S(O)₂—O—, or(R_(d))₂P(O)—O— (R_(d) is H, OH, alkyl, alkoxy, amino, or aryl; e.g.,R_(d) is ethyl); R^(2′) is R—O— (R is alkyl substituted with amino), orR^(2′) is R_(d)S(O)₂—O—, or (R_(a))₂P(O)—O— (R_(d) is H, OH, alkyl,alkoxy, amino, or aryl, e.g., R_(d) is ethyl); and one of R^(1′),R^(3′), and R^(4′) is R_(d)S(O)₂—O—, or (R_(d))₂P(O)—O— (R_(d) is H, OH,alkyl, alkoxy, amino, or aryl, e.g., R_(d) is ethyl).

Still another aspect of this invention relates to a compound of Formula(III):

wherein each of X is N or CH; each of R¹, R², R³, R⁴, R⁵, R^(1′),R^(2′), R^(3′), R^(4′), and R^(5′), independently, is H, alkyl (e.g.,alky substituted with halo or SO₂R_(d)), halo, OH, R_(c)—O—,R_(d)S(O)₂—O—, or (R_(d))₂P(O)—O—, R_(c) being unsubstituted alkyl oralkyl substituted with halo, OH, alkoxy, amino, cycloalkyl,heterocycloalkyl, aryl, or heteroaryl, and R_(d) being H, OH, alkyl,alkoxy, amino, or aryl; and R⁶ is C(O)—R_(e), C(O)NR_(e)R_(f), or alkyl(e.g., unsubstituted alkyl or alkyl substituted with cycloalkyl), eachof R_(e) and R_(f), independently, being alkyl or cycloalkyl.

Referring to Formula (III), a subset of compounds have one or more ofthe following features: X is N; R² is OH or R—O— (R is alkyl substitutedwith amino, e.g., R is CH₂H₂N(C₂H₅)₂); and R⁶ is cyclopropylcarbonyl orcyclopropylmethyl.

The term “alkyl” refers to a saturated, linear or branched hydrocarbonmoiety containing 1-10 carbon atoms, such as —CH₃ or —CH(CH₃)₂. The term“cycloalkyl” refers to a 3-10 membered, saturated, cyclic hydrocarbonmoiety, such as cyclohexyl. The term “heterocycloalkyl” refers to a 3-10membered, saturated, cyclic moiety having at least one ring heteroatom(e.g., N, O, or S), such as 4-tetrahydropyranyl. The term “aryl” refersto a hydrocarbon moiety having one or more aromatic rings. Examples ofaryl moieties include phenyl (Ph), phenylene, naphthyl, naphthylene,pyrenyl, anthryl, and phenanthryl. The term “heteroaryl” refers to amoiety having one or more aromatic rings that contain at least oneheteroatom (e.g., N, O, or S). Examples of heteroaryl moieties includefuryl, furylene, fluorenyl, pyrrolyl, thienyl, oxazolyl, imidazolyl,thiazolyl, pyridyl, pyrimidinyl, quinazolinyl, quinolyl, isoquinolyl andindolyl.

Alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl mentionedherein include both substituted and unsubstituted moieties, unlessspecified otherwise. Possible substituents on cycloalkyl,heterocycloalkyl, aryl, and heteroaryl include, but are not limited to,C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀cycloalkenyl, C₁-C₂₀ heterocycloalkyl, C₁-C₂₀ heterocycloalkenyl, C₁-C₁₀alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, amino, C₁-C₁₀alkylamino, C₁-C₂₀ dialkylamino, arylamino, diarylamino,alkylsulfonamino, arylsulfonamino, C₁-C₁₀ alkylimino, arylimino, C₁-C₁₀alkylsulfonimino, arylsulfonimino, hydroxyl, halo, thio, C₁-C₁₀alkylthio, arylthio, C₁-C₁₀ alkylsulfonyl, arylsulfonyl, acylamino,aminoacyl, aminothioacyl, amidino, guanidine, ureido, cyano, nitro,nitroso, azido, acyl, thioacyl, acyloxy, carboxyl, and carboxylic ester.On the other hand, possible substituents on alkyl include all of theabove-recited substituents except C₁-C₁₀ alkyl. Cycloalkyl,heterocycloalkyl, aryl, and heteroaryl can also be fused with eachother.

The compounds described above include the compounds themselves, as wellas their salts, prodrugs, and solvates, if applicable. A salt, forexample, can be formed between an anion and a positively charged group(e.g., ammonium ion) on a 1,5-diphenyl-penta-1,4-dien-3-one compound.Suitable anions include chloride, bromide, iodide, sulfate, nitrate,phosphate, citrate, methanesulfonate, trifluoroacetate, acetate,succinate, malate, tosylate, tartrate, fumurate, glutamate, glucuronate,lactate, glutarate, and maleate. Likewise, a salt can also be formedbetween a cation and a negatively charged group (e.g., carboxylate) on a1,5-diphenyl-penta-1,4-dien-3-one compound. Suitable cations includesodium ion, potassium ion, magnesium ion, calcium ion, and an ammoniumcation. The compounds may also be in prodrug and solvate form. Examplesof prodrugs include esters and other pharmaceutically acceptablederivatives, which, upon administration to a subject, are capable ofproviding active compounds. A solvate refers to a complex formed betweenan active compound and a pharmaceutically acceptable solvent. Examplesof pharmaceutically acceptable solvents include water, ethanol,isopropanol, ethyl acetate, acetic acid, and ethanolamine.

The compounds contain non-aromatic double bonds. Thus, they can occur ascis- or trans-isomeric forms. Such isomeric forms are contemplated.

Still another aspect of this invention relates to a method for treatingcancer, inflammatory disease, or autoimmune disease. The method includesadministering to a subject in need thereof an effective amount of one ormore of the above described compounds.

Also within the scope of this invention is a composition containing oneor more of the compounds described above and a pharmaceuticallyacceptable carrier for use in treating cancer/inflammatorydisease/autoimmune disease, and the use of such a composition for themanufacture of a medicament for the just-mentioned treatment.

The details of one or more embodiments of the invention are set forth inthe description below. Other features, objects, and advantages of theinvention will be apparent from the description and from the claims.

DETAILED DESCRIPTION

The compounds described above can be prepared by methods well known inthe art. Schemes 1, 2, and 3 below demonstrate general synthetic routesused to synthesize compounds of formulas (I), (II), and (III),respectively.

As shown in Scheme 1 above, benzaldehyde is condensed with excessiveacetone under a basic condition to give a 4-phenylbut-3-en-2-onecompound, which is condensed with a second benzaldehyde compound toafford compounds of Formula (I). The thus-obtained product can befurther modified to prepare other compounds of Formula (I).

As shown in Scheme 2 above, hydroxyl-substituted1,5-diphenylpenta-1,4-dien-3-one compounds (which can be prepared by themethod described above) are condensed with sulfonyl chloride orchlorophosphine/phosphorochloridate/phosphoryl trichloride or acylchloride to give compounds of Formula (II). Similarly,3,5-dibenzylidenepiperidin-4-one compounds can be condensed withsulphonyl chloride/chlorophosphine/acyl chloride to give compounds alsocovered by Formula (II).

As shown in Scheme 3 above, piperidin-4-one is reacted with acylchloride or bromoalkane to give N-substituted piperidin-4-one, which issubsequently condensed with 2 equivalents of benzaldehyde to affordsymmetric 3,5-dibenzylidenepiperidin-4-one compounds covered by Formula(III). The N-substituted piperidin-4-one can be first reacted with 1equivalent of benzaldehyde, and then with 1 equivalent of secondbenzaldehyde to prepare asymmetric 3,5-dibenzylidenepiperidin-4-onecompounds, which are also covered by Formula (III).

Shown below are exemplary compounds of Formula (I), (II), and (III)prepared by the above-described methods:

Compound Nos. Structures  1

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The compounds described above deactivate STAT proteins. Thus, thisinvention covers a method of administering an effective amount of one ormore of the compounds to a patient having cancer, inflammatory disease,or autoimmune disease.

The term “treating” or “treatment” refers to administering one or morecompounds to a subject, who has an above-described condition, a symptomof such a condition, or a predisposition toward such a condition, withthe purpose to confer a therapeutic effect, e.g., to cure, relieve,alter, affect, ameliorate, or prevent the above-described disorder, thesymptom of it, or the predisposition toward it. “An effective amount”refers to the amount of an active compound that is required to confer atherapeutic effect on the treated subject. Effective doses will vary, asrecognized by those skilled in the art, depending on the types ofdiseases treated, route of administration, excipient usage, and thepossibility of co-usage with other therapeutic treatment.

Cancers that can be treated by the method of the invention include bothsolid and haematological tumors of various organs. Examples of solidtumors include pancreatic cancer, bladder cancer, colon cancer,colorectal cancer, breast cancer (e.g., metastatic breast cancer),prostate cancer (e.g., androgen-dependent, androgen-independent, orcastrate-resistant prostate cancer), renal cancer (e.g., metastaticrenal cell carcinoma), hepatocellular cancer, lung cancer (e.g.,non-small cell lung cancer, bronchioloalveolar carcinoma, oradenocarcinoma of the lung), ovarian cancer (e.g., progressiveepithelial or primary peritoneal cancer), cervical cancer, gastriccancer, esophageal cancer, head and neck cancer (e.g., squamous cellcarcinoma of the head and neck), melanoma, neuroendocrine cancer (e.g.,metastatic neuroendocrine tumors), brain tumors (e.g., glioma,anaplastic oligodendroglioma, adult glioblastoma multiforme, or adultanaplastic astrocytoma), bone cancer, and soft tissue sarcoma. Examplesof haematological tumors include various leukemia (e.g., myeloidleukemia, chronic myelogenous leukemia or CML [accelerated CML and CMLblast phase], acute lymphoblastic leukemia, or chronic lymphocyticleukemia), Hodgkin's disease, non-Hodgkin's lymphoma (e.g., follicularlymphoma or mantle cell lymphoma), B-cell lymphoma, T-cell lymphoma,multiple myeloma, Waldenstrom's macroglobulinemia, myelodysplasticsyndromes (e.g., refractory anemia, refractory anemia with ringedsiderblasts, refractory anemia with excess blasts or RAEB, or RAEB intransformation), and myeloproliferative syndromes.

Inflammatory diseases that can be treated by the method of thisinvention include, but are not limited to, asthma, atherosclerosis,rheumatoid arthritis, inflammatory bowel diseases, Crohn, ulcerativecolitis, ischemic heart disease, cardiomyopathy, glomerulonephritis,nephritic syndrome, hepatitis B or C infection, respiratory syncytialvirus infection (pulmonary), and Guillain-Barré syndrome.

Autoimmune diseases that can be treated by the method of this inventioninclude, but are not limited to, allergic encephalopathy, chronicobstructive pulmonary disease, psoriasis, psoriatic arthritis, diabetesmellitus, systemic lupus erythematosus, multiple sclerosis,polymyositis, dermatomyositis, mixed connective tissue disease,Sjögren's syndrome, Wegener's granulomatosis, polyarteritis nodosa,rheumatoid arthritis, and idiopathic thrombocytopenic purpura.

To practice the method of the present invention, a composition havingone or more of the above-described compounds can be administeredparenterally, orally, nasally, rectally, topically, or buccally. Theterm “parenteral” as used herein refers to subcutaneous, intracutaneous,intravenous, intramuscular, intraarticular, intraarterial,intrasynovial, intrasternal, intrathecal, intralesional, or intracranialinjection, as well as any suitable infusion technique.

A sterile injectable composition can be a solution or suspension in anon-toxic parenterally acceptable diluent or solvent, such as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that canbe employed are mannitol, water, Ringer's solution, and isotonic sodiumchloride solution. In addition, fixed oils are conventionally employedas a solvent or suspending medium (e.g., synthetic mono- ordiglycerides). Fatty acid, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions can also contain a long chain alcohol diluent or dispersant,carboxymethyl cellulose, or similar dispersing agents. Other commonlyused surfactants such as Tweens or Spans or other similar emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms can also be used for the purpose of formulation.

A composition for oral administration can be any orally acceptabledosage form including capsules, tablets, emulsions and aqueoussuspensions, dispersions, and solutions. In the case of tablets,commonly used carriers include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions or emulsions areadministered orally, the active ingredient can be suspended or dissolvedin an oily phase combined with emulsifying or suspending agents. Ifdesired, certain sweetening, flavoring, or coloring agents can be added.

A nasal aerosol or inhalation composition can be prepared according totechniques well known in the art of pharmaceutical formulation. Forexample, such a composition can be prepared as a solution in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or othersolubilizing or dispersing agents known in the art.

A composition having one or more active compounds can also beadministered in the form of suppositories for rectal administration.

The carrier in the pharmaceutical composition must be “acceptable” inthe sense that it is compatible with the active ingredient of thecomposition (and preferably, capable of stabilizing the activeingredient) and not deleterious to the subject to be treated. One ormore solubilizing agents can be utilized as pharmaceutical excipientsfor delivery of an active 1,5-diphenyl-penta-1,4-dien-3-one compound.Examples of other carriers include colloidal silicon oxide, magnesiumstearate, cellulose, sodium lauryl sulfate, and D&C Yellow #10.

The 1,5-diphenyl-penta-1,4-dien-3-one compounds described above can bepreliminarily screened for their efficacy in deactivating STAT proteinsand treating above-described diseases by an in vitro assay and thenconfirmed by animal experiments and clinic trials. Other methods willalso be apparent to those of ordinary skill in the art.

The specific examples below are to be construed as merely illustrative,and not limitative of the remainder of the disclosure in any waywhatsoever. Without further elaboration, it is believed that one skilledin the art can, based on the description herein, utilize the presentinvention to its fullest extent. All publications cited herein arehereby incorporated by reference in their entirety.

Chemical Syntheses

Melting points were determined using a Fisher-John melting pointapparatus without calibration. Proton Nuclear Magnetic Resonance (¹HNMR) and ¹³C NMR spectra were measured on Varian Gemini 300 or Inova 400spectrometers with tetramethylsilane as the internal standard. Chemicalshifts were reported in δ (ppm). Mass spectra (MS) were obtained on aShimadzu LCMS-2010. A CombiFlash chromatographic system was performedover Grace silica gel cartridge for general separation and purification.Preparative thin layer chromatography using silica gel plates (Kieselgel60, F254, 1.00 mm) were also used for separation and purification.Precoated silica gel plates (Kieselgel 60, F254, 0.25 mm) were used forthin layer chromatography (TLC) analysis. All reagents and solvents werepurchased from Aldrich, Fisher, VWR, or other venders.

Synthesis of Compounds 1-4 and 21

Compounds 1-4 and 21 were prepared as shown in Scheme 4 below:

To a solution of hydroxyl benzaldehyde in DMF was added K₂CO₃ (2 eq.each hydroxyl group) at 4° C. in an ice-bath and methyl chloromethylether (MOM chloride) (1.3 eq. each hydroxyl group). After the solutionwas stirred at room temperature and monitored by TLC for 3-5 hours,hexanes/dichloromethane (1:1) was added and allowed to stir for 30 min.The solid was filtered out and the filtrate was concentrated. Theresulting residue was diluted with EtOAc and washed with H₂O twice. Theaqueous layer was extracted with EtOAc. The combined organic layers weredried over Na₂SO₄ and concentrated. The crude was purified by aCombiFlash chromatography system using a Grace silica gel column andhexanes/ethyl acetate (as the eluent) to give MOM protectedhydroxylbenzaldehyde. The resulting compound further reacted with excessacetone in ethanol catalyzed by 10% NaOH (1.2 eq. of NaOH). Afterstirred at room temperature and monitored by TLC for 1-5 hours, thereaction mixture was diluted with CH₂Cl₂, washed with H₂O twice, andextracted with CH₂Cl₂ twice. The MOM protectedhydroxy-phenyl-but-3-en-2-one was obtained after purification by aCombiFlash system using hexanes/ethyl acetate as the eluent. Theresulting compound reacted with an appropriate MOM protectedhydroxybenaldehyde to afford MOM protected hydroxyl1,5-diphenyl-penta-1,4-dien-3-one, which was deprotected by heating in50% acetic acid aqueous solution to afford the desired product.

Compound 1: yellow crystalline solid; ESI MS m/z: 251.10 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.75 (d, 1H, J=15.0 Hz, H-1), 7.63 (d, 1H, J=15.0Hz, H-5), 7.64 (m, 2H, aromatic ring H), 7.43-7.41 (m, 2H, aromatic ringH), 7.72-7.27 (m, 2H, aromatic ring H), 7.21-7.19 (d, 1H, J=6.0 Hz,aromatic ring H), 7.13-7.12 (m, 1H, aromatic ring H), 7.08 (d, 1H,J=15.0 Hz, H-2), 7.06 (d, 1H, J=15.0 Hz, H-4), 6.92-6.83 (m, 1H,aromatic ring H), 5.38 (br. 1H, OH).

Compound 2: yellow crystalline solid; ESI MS m/z: 267.29 [M+H]⁺; ¹H NMR(300 MHz, CD₃OD₃) δ: 8.10 (d, 1H, J=15.0 Hz, H-1), 7.63 (d, 1H, J=15.0Hz, H-5), 7.75-7.62 (m, 1H, aromatic ring H), 7.25 (d, 1H, J=15.0 Hz,H-4), 7.22 (d, 1H, J=15.0 Hz, H-2), 7.21-7.11 (m, 4H, aromatic ring H),6.91-6.84 (m, 3H, aromatic ring H), 3.68 (br. 2H, OH).

Compound 3: yellow crystalline solid; ESI MS m/z: 281.05 [M−H]⁻; ¹H NMR(300 MHz, CD₃OD₃) δ: 7.68 (d, 1H, J=15.9 Hz, H-5), 7.66 (d, 1H, J=15.9Hz, H-1), 7.27-6.97 (m, 5H, aromatic ring H), 6.99 (d, 1H, J=15.9 Hz,H-4), 6.88-6.84 (m, 2H, aromatic ring H), 6.80 (d, 1H, J=15.9 Hz, H-2),4.91 (br. 3H, OH).

Compound 4: yellow crystalline solid; ESI MS m/z: 281.05 [M−H]⁻; ¹H NMR(300 MHz, CD₃OD₃) δ: 7.70 (d, 1H, J=15.9 Hz, H-5), 7.65 (d, 1H, J=15.9Hz, H-1), 7.60-7.56 (dd, 2H, aromatic ring H), 7.15-7.06 (m, 2H,aromatic ring H), 7.12 (d, 1H, J=15.9 Hz, H-4), 6.98 (d, 1H, J=15.9 Hz,H-2), 6.81 (t, 3H, J=8.4 Hz, aromatic ring H).

Compound 21: yellow crystalline solid; ESI MS m/z: 281.09 [M−H]⁻; ¹H NMR(300 MHz, CD₃OD₃) δ: 7.63 (d, 1H, J=15.9 Hz, H-5), 7.56 (d, 1H, J=15.9Hz, H-1), 7.21-7.05 (m, 3H, aromatic ring H), 7.12 (d, 1H, J=15.9 Hz,H-4), 7.03 (d, 1H, J=15.9 Hz, H-2), 6.82-6.778 (m, 1H, aromatic ring H),6.57 (d, 2H, J=2.1 Hz, aromatic ring H), 6.29 (t, 1H, J=2.1 Hz, aromaticring H).

Syntheses of Compounds 20, 30, 31, 38, 39, 41, 42, 44, 62-64, 73, and 75

Compound 20, 30, 31, 38, 39, 41, 42, 44, 62-64, 73, and 75 were preparedas shown in Scheme 5 below:

Methoxy substituted benzaldehyde was reacted with acetone followed bycondensation with second methoxy substituted benzaldehyde in the samemanner as described above to afford methoxy substituted1,5-diphenyl-penta-1,4-dien-3-ones compounds 30 and 31 or methyl methoxysubstituted 1,5-diphenyl-penta-1,4-dien-3-ones. Demethylation (orpartial demethylation) with BBr₃ (2 eq. each methoxy group) in CH₂Cl₂from −78° C. to 0° C. to room temperature gave the desired phenoliccrude products. The reaction was monitored by TLC. After completion, thereaction mixture was poured into acidic ice/water and then extracted byethyl ether. The desired products (i.e., compounds 20, 62-64) wereobtained after purification over a CombiFlash chromatography system.

Compound 20: red crystalline solid; ESI MS m/z: 281.10 [M−H]⁻; ¹H NMR(300 MHz, CD₃OD₃) δ: 8.06 (d, 1H, J=15.9 Hz, H-5), 7.63 (d, 1H, J=15.9Hz, H-1), 7.26-7.05 (m, 5H, aromatic ring H and H-2,4), 6.86-6.62 (m,4H, aromatic ring H).

Compound 62: Amorphous, ESI MS m/z: 309.0 [M+H]⁺; ¹H NMR (400 MHz,acetone-d₆) δ: 8.03-7.94 (m, 2H, H-1,5), 7.15-7.00 (m, 3H, aromatic ringH), 6.96-6.87 (m, 3H, aromatic ring H), 6.68 (d, J=16.0 Hz, 1H, H-2),6.65 (d, J=16.0 Hz, 1H, H-4), 3.85 (s, 3H, OCH₃), 2.12 (s, 3H, CH₃),2.10 (s, 3H, CH₃).

Compound 63: Bright yellow solid, ESI MS m/z: 295.1 [M+H]⁺; ¹H NMR (400MHz, CD₃OD) δ: 8.01 (d, 2H, J=15.6 Hz, H-1,5), 7.66-7.64 (m, 2H,aromatic ring H), 6.98 (d, 2H, J=15.6 Hz, H-2,4), 6.66-6.64 (m, 4H,aromatic ring H),), 2.40 (s, 6H, CH₃).

Compound 64: Amorphous, ESI MS m/z: 309.1 [M+H]⁺; ¹H NMR (400 MHz,acetone-d₆) δ: 8.07-7.94 (m, 2H, H-1,5), 7.75-7.67 and 7.08-7.01 (m, 1H,aromatic ring H), 7.50-7.44 (m, 2H, aromatic ring H), 6.83-6.73 (m, 1H,aromatic ring H), 6.70-6.66 (m, 2H, aromatic ring H),), 6.48 (d, J=16.0Hz, 1H, H-2), 6.47 (d, J=16.0 Hz, 1H, H-4), 3.81 (s, 3H, OCH₃), 1.98 (s,3H, CH₃), 1.94 (s, 3H, CH₃).

Compounds 38, 39, 41, 42, 44, 73, and 75 were synthesized by reaction ofsubstituted benzaldehyde with (E)-4-(3-hydroxyphenyl)but-3-en-2-one inethanol/20% NaOH aqueous solution at room temperature as shown in Scheme5 above. The reactions were monitored by TLC. After completion, thesolutions were neutralized by acetic acid, extracted with ethyl acetate,and concentrated to afford the desired products. To furnish Compound 44,the THP protecting group, which was introduced to make3-methoxy-4-(tetrahydro-pyran-2-yloxy)-benzaldehyde, was removed bytreating with 0.1 eq. pyridinium p-toluenesulfonate in ethanol solutionat room temperature.

Compound 38: pale yellow needles; mp 159-160° C.; C₁₉H₁₈O₄, ESI-MS: m/z311.2 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 7.75, 7.65 (1H each, both d,J=16.2 Hz, H-1,5), 7.42 (1H, d, J=2.1 Hz, H-2′), 7.34 (1H, dd, J=2.1,8.4 Hz, H-6′), 7.25, 7.23 (1H each, both d, J=16.2 Hz, H-2,4), 7.28-7.13(2H, m, H-4″, 5″), 7.14 (1H, br, t, H-2″), 7.04 (1H, d, J=8.4 Hz, H-5′),6.86 (1H, br. d, 8.4 Hz, H-6″), 3.84, 3.82 (both s, 3H each, OCH₃×2).

Compound 39: yellow fine crystals; mp 152-153° C.; C₁₈H₁₆O₃, ESI-MS: m/z281.2 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 7.76, 7.62 (1H each, both d,J=16.2 Hz, H-1,5), 7.72 (2H, d, J=9.0 Hz, H-2′, 6′), 7.22 (1H, d, 8.0Hz, H-6″), 7.18, 7.17 (1H each, both d, J=16.2 Hz, H-2,4), 7.17 (1H, m,5″), 7.09 (1H, br. 3, H-2″), 6.98 (2H, d, J=9.0 Hz, H-3′, 5′), 6.81 (1H,br. d, 8.0 Hz, H-4″), 3.77 (s, 3H, OCH₃).

Compound 41: pale yellow needles; mp 138-139° C.; C₁₈H₁₃F₃O₂, ESI-MS:m/z 319.2 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 8.14 (1H, br., H-2′), 8.07(1H, br. d, J=7.8 Hz, H-6′), 7.81, 7.50 (1H each, both d, J=15.9 Hz,H-1, 2), 7.78-7.62 (2H, m, H-4′, 5′), 7.69, 7.19 (1H each, both d,J=16.2 Hz, H-4, 5), 7.22 (2H, 4″, 5″), 7.10 (1H, br. s, H-2″), 6.83 (1H,br. d, J=8.7 Hz, H-6″).

Compound 42: pale yellow fine crystals; mp 159-160° C.; C₁₇H₁₃FO₂,ESI-MS: m/z 269.2 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 7.92 (1H, dt,J=1.8, 7 Hz, H-4′), 7.73, 7.42 (1H each, both d, J=16.2 Hz, H-1, 2),7.69, 7.16 (1H each, both d, J=16.2, Hz, H-4, 5), 7.47 (1H, m, H-5-″),7.30-7.12 (3H, m, 2′, 5′, 6′, 4″), 7.10 (1H, br. s, H-2″), 6.82 (1H, dt,J=1.8, 7.2 Hz, H-6″).

Compound 44: yellow fine crystals; mp 144-145° C.; C₂₃H₂₆O₁₀S₃, ESI-MS:m/z 559.1 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 7.63-6.93 (11H, m, aromaticH and vinyl H), 3.81 (2H, s, OCH₃).

Compound 73: Off-white solid, ESI MS m/z: 269.5 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ: 7.68 (d, J=16.0 Hz, 1H, H-1), 7.67 (d, J=16.0 Hz, 1H, H-5),7.37-7.35 (m, 2H, aromatic ring H), 7.29-7.24 (m, 2H, aromatic ring H),7.18-7.16 (m, 1H, aromatic ring H), 7.11-7.08 (m, 2H, aromatic ring H)),7.04 (d, J=16.0 Hz, 1H, H-4), 7.01 (d, J=16.0 Hz, 1H, H-2), 6.91-6.88(m, 1H, aromatic ring H).

Compound 75: Light yellow solid, ESI MS m/z: 335.5 [M+H]⁺; ¹H NMR (400MHz, CDCl₃) δ: 7.69 (d, J=15.6 Hz, 1H, H-1), 7.67 (d, J=15.6 Hz, 1H,H-5), 7.49-7.47 (m, 1H, aromatic ring H), 7.42-7.39 (m, 2H, aromaticring H), 7.28-7.22 (m, 2H, aromatic ring H), 7.16-7.14 (m, 2H, aromaticring H),), 7.05 (d, J=16.0 Hz, 1H, H-4), 7.02 (d, J=16.0 Hz, 1H, H-2),6.95-6.92 (m, 1H, aromatic ring H).

Syntheses of Compounds 12, 13, and 15-17

Compounds 12, 13, and 15-17 were synthesized as shown in Scheme 6, bymethods modified based on that described in Roberta Costi, et al.,Bioorganic & Medicinal Chemistry 2004, 12: 199-215.

To a solution of piperidin-4-one in DMF was added triethylamine (1.5eq.) and N,N-diethylacetamide chloride (1.5 eq.). The resulting solutionwas stirred at room temperature for 5 hours or until completion observedby TLC. The solvent was evaporated and the residue was partitioned inH₂O and EtOAc. The aqueous layer was extracted with EtOAc and thecombined organic layers were dried over Na₂SO₄ and concentrated. Thecrude product was purified by a CombiFlash chromatography system usingsilica gel cartridge and CH₂Cl₂/MeOH gradient eluent to afford4-oxo-piperidine-1-carboxylic acid diethylamide, which was subsequentlyreacted with 3-hydroxybenzaldehyde (2.5 eq.) in acetic acid (99.7%) andpurged with HCl gas (0.5-1 h). After stirring at room temperature for 3hours, the solvent was evaporated and the crude was purified by aCombiflash chromatography system with hexanes/EtOAc as eluent, followedby crystallization from MeOH, to afford Compound 12 as a yellowcrystalline solid.

Compound 12: yellow crystalline solid; ESI MS m/z: 407.49 [M+H]⁺; ¹H NMR(300 MHz, CD₃OD) δ: 7.61 (s. 2H, benzylidene CH═), 7.16 (t, 2H, J=7.5Hz, aromatic ring H), 6.82-6.79 (m, 2H, aromatic ring H), 6.75-6.71 (m,4H, aromatic ring H), 4.36 (s, 4H, 4-oxo-piperidine-H-2, 6), 2.95 (q,4H, —NCH₂ CH₃), 0.71 (t, 6H, —NCH₂CH₃ ).

Compound 16 was prepared in the same manner as described above exceptthat cyclopropanecarbonyl chloride (1.5 eq.) was used instead ofN,N-diethylacetamide chloride.

Compound 16: yellow solid; ESI MS m/z: 376.16 [M+H]⁺; ¹H NMR (300 MHz,DMSO-d₆) δ: 7.59 (s. 2H, benzylidene CH═), 7.30 (t, 2H, J=7.5 Hz,aromatic ring H), 6.98-6.93 (m, 4H, aromatic ring H), 6.87-6.85 (m, 2H,aromatic ring H), 3.39 (s, 4H, 4-oxo-piperidine-H-2, 6), 1.69 (m, 1H,cyclopropanecarbonyl CH), 0.62-0.58 (m, 4H, cyclopropanecarbonyl CH₂ ).

Compounds 15 and 17 were synthesized by reaction of piperidin-4-one with1-bromo-butane (1.5 eq.) or bromo methyl-cyclopropane (1.5 eq.) in DMFin the presence of K₂CO₃. After stirring at room temperature for 24 h ormonitored by MS, the reaction solvent was removed by evaporation. Theresidue was partitioned in H₂O and EtOAc, and the organic layer waswashed with H₂O twice. The aqueous then was extracted with EtOAc and theextract was dried over Na₂SO₄. The crude product was purified by aCombiflash chromatography system using silica gel cartridge andCH₂Cl₂/MeOH gradient eluent. The obtained N-substituted 4-oxo-piperidinecompound was condensed with 3-hydroxybenzaldehyde (2.5 eq.), followingthe same procedure described above, to afford the desired product.

Compound 15: yellow solid; ESI MS m/z: 364.19 [M+H]⁺; ¹H NMR (300 MHz,CD₃OD) δ: 7.64 (s. 2H, benzylidene CH═), 7.21 (t, 2H, J=8.1 Hz, aromaticring H), 6.85 (d, br, 2H, J=8.1 Hz, aromatic ring H), 6.80-6.76 (m, 4H,aromatic ring H), 3.80 (s, 4H, 4-oxo-piperidine-H-2, 6), 2.50 (t, 2H,J=8.1 Hz, —NCH₂ CH₂—), 1.42-1.30 (m, 2H, —NCH₂CH₂ CH₂—), 1.27-1.14 (m,2H, —NCH₂CH₂CH₂ CH₃), 0.81 (t, 3H, —NCH₂CH₂CH₂CH₃ ).

Compound 17: yellow solid; ESI MS m/z: 362.18 [M+H]⁺; ¹H NMR (300 MHz,DMSO-d₃) δ: 7.66 (s. 2H, benzylidene CH═), 7.21 (t, 2H, J=7.8 Hz,aromatic ring H), 6.86 (d, br, 2H, J=7.2 Hz, aromatic ring H), 6.81-6.75(m, 4H, aromatic ring H), 3.90 (s, 4H, 4-oxo-piperidine-H-2, 6), 2.40(d, 2H, J=6.6 Hz, methylene-cyclopropane-CH₂ ), 0.65 (m, 1H,cyclopropane CH), 0.43-0.37 (m, 2H, cyclopropane CH₂ ), 0.11-0.07 (m,2H, cyclopropane CH₂ ).

Compound 13 was synthesized starting from treatingoxo-(4-oxo-cyclohexyl)-acetic acid with1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (2 eq.) and4-dimethylaminopyridine (catalytic amount) in DMF. After stirring atroom temperature for approximately 10 min., the reaction mixture wascooled in an ice-bath and diethylamine (1.5 eq.) was added. Theresulting mixture was stirred at room temperature for 5 hours with TLCmonitoring. The solvent was removed by evaporation. The residue wasdiluted with EtOAc and washed with H₂O twice. The aqueous then wasextracted with EtOAc and the organic layer was dried over Na₂SO₄. Thedesired intermediate N,N-diethyl-2-oxo-2-(4-oxo-cyclohexyl)-acetamidewas obtained after purification through a combiflash chromatographysystem using a silica gel cartridge and CH₂Cl₂/MeOH gradient eluent,which further condensed with 3-hydroxybenzaldehyde (2.5 eq.) byfollowing the same procedure described above to get the desired productas a yellow solid ESI MS m/z: 406.51 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆)δ: 7.54 (s. 2H, benzylidene CH═), 7.25 (t, 2H, J=7.8 Hz, aromatic ringH), 6.93-6.89 (m, 4H, aromatic ring H), 6.81-6.78 (m, 2H, aromatic ringH), 3.23 (q, 4H, J=7.2 Hz, —NCH₂ CH₃), 2.95 (m, 4H,4-oxo-cyclohexyl-H-2,6), 2.54 (m, 1H, cyclohexyl-H-1), 0.94 (m, 6H,—NCH₂CH₃ ).

Syntheses of Compounds 5-11, 14, 18, 22-29, 32-34, 37, 40, 43, 53,65-66, 69-70, 74, 77, and 78

Syntheses of these compounds are shown in Scheme 7.

To a solution of 1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one(synthesized by the method shown in Scheme 4) in CH₂Cl₂ (containing asmall amount of DMF) was slowly added ethanesulfonyl chloride (˜10 eq.)and Et₃N (˜10 eq.). The resulting mixture was stirred at roomtemperature for 4-5 hours. The reaction mixture was diluted with CH₂Cl₂,washed with water twice, and extracted with CH₂Cl₂. The organic extractwas dried over Na₂SO₄, filtered, and concentrated to get crude as ayellow solid. The crude was purified by quick column filtration, thencrystallization, and re-crystallization from EtOAc to get the desiredproduct compound 6 as a light yellow crystalline solid. Yield>78%. ESIMS m/z: 451.2 [M+H]⁺; ¹H NMR (300 MHz, CDCl₃) δ: 7.71 (d, 2H, J=15.9 Hz,H-1,5), 7.57-7.53 (m, 4H, aromatic ring H), 7.47 (t, 2H, J=7.8 Hz,aromatic ring H), 7.34-7.31 (m, 2H, aromatic ring H), 7.08 (d, 2H,J=15.9 Hz, H-2,4), 3.34 (q, 4H, J=7.5 Hz, —OSO₂CH₂ CH₃), 1.58 (t, 6H,J=7.5 Hz, —OSO₂CH₂CH₃ ).

Compounds 5, 7-11, 22, 26, 27, 40, 43, 53, 65-66, 69-70, 74, and 77 weresynthesized in the manner similar to that described above.

Compound 5: a light yellow crystalline solid; ESI MS m/z: 451.2 [M+H]⁺;¹H NMR (300 MHz, CDCl₃) δ: 7.72 (d, 2H, J=15.9 Hz, H-1,5), 7.66 (d, 4H,J=8.7 Hz, aromatic ring H), 7.34 (d, 4H, J=8.7 Hz, aromatic ring H),7.04 (d, 2H, J=15.9 Hz, H-2,4), 3.33 (q, 4H, J=7.5 Hz, —OSO₂CH₂ CH₃),1.57 (t, 6H, J=7.5 Hz, —OSO₂CH₂CH₃ ).

Compound 7: yellow crystalline solid; ESI MS m/z: 343.4 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.76 (d, 1H, J=15.9 Hz, H-5), 7.70 (d, 1H, J=15.9Hz, H-1), 7.65-7.62 (m, 2H, aromatic ring H), 7.57-7.53 (m, 2H, aromaticring H), 7.49-7.42 (m, 4H, aromatic ring H), 7.34-7.30 (m, 1H, aromaticring H), 7.10 (d, 1H, J=15.9 Hz, H-4), 7.09 (d, 1H, J=15.9 Hz, H-2),3.33 (q, 2H, J=7.5 Hz, —OSO₂CH₂ CH₃), 1.58 (t, 3H, J=7.5 Hz, —OSO₂CH₂CH₃).

Compound 8: off-white crystalline solid; ESI MS m/z: 423.2 [M+H]⁺; ¹HNMR (300 MHz, CDCl₃) δ: 7.70 (d, 2H, J=15.9 Hz, H-1,5), 7.58-7.38 (m,4H, aromatic ring H), 7.48 (t, 2H, J=7.5 Hz, aromatic ring H), 7.36-7.33(m, 2H, aromatic ring H), 7.08 (d, 2H, J=15.9 Hz, H-2,4), 3.21 (s, 6H,—OSO₂CH₃ ).

Compound 9: yellow crystalline solid; ESI MS m/z: 479.2 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.71 (d, 2H, J=15.9 Hz, H-1,5), 7.57-7.53 (m, 4H,aromatic ring H), 7.47 (t, 2H, J=7.8 Hz, aromatic ring H), 7.34-7.31 (m,2H, aromatic ring H), 7.08 (d, 2H, J=15.9 Hz, H-2,4), 3.31-3.26 (m, 4H,—OSO₂CH₂ CH₂CH₃), 2.12-1.99 (m, 4H, —OSO₂CH₂CH₂ CH₃), 1.16 (t, 6H, J=7.5Hz, —OSO₂CH₂CH₂ CH₃).

Compound 10: light yellow thick oil; ESI MS m/z: 547.2 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.88-7.85 (m, 4H, phenyl sulfonate aromatic ring H),7.71 (t, 2H, phenyl sulfonate aromatic ring H), 7.59 (d, 2H, J=15.9 Hz,H-1,5), 7.59-7.48 (m, 6H, phenyl sulfonate substituted and biphenylaromatic ring H), 7.35 (t, 2H, J=7.8 Hz, aromatic ring H), 7.27-7.25 (m,2H, biphenyl aromatic ring H), 7.03-7.00 (m, 2H, biphenyl aromatic ringH), 6.95 (d, 2H, J=15.9 Hz, H-2,4).

Compound 11: a light yellow crystalline solid; ESI MS m/z: 559.1 [M+H]⁺;¹H NMR (300 MHz, CDCl₃) δ: 7.73 (d, 1H, J=15.9 Hz, H-5), 7.72-7.70 (dd,1H, aromatic ring H), 7.67 (d, 1H, J=15.9 Hz, H-1), 7.67 (d, 2H, J=8.7Hz, 2″,6″aromatic ring H), 7.55-7.50 (m, 2H, aromatic ring H), 7.34 (d,2H, J=8.7 Hz, 3″,5″aromatic ring H), 7.05 (d, 1H, J=15.9 Hz, H-4), 7.03(d, 1H, J=15.9 Hz, H-2), 3.47 (m, 6H, —OSO₂CH₂ CH₃), 1.58 (m, 9H,—OSO₂CH₂CH₃ ).

Compound 26: amorphous; ESI MS m/z: 559.1 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 8.02 (d, 1H, J=15.9 Hz, H-1), 7.72-7.70 (dd, 1H, aromatic ringH), 7.73 (d, 1H, J=15.9 Hz, H-5), 7.59-7.33 (m, 6H, aromatic ring H),7.20 (d, 1H, J=15.9 Hz, H-2), 7.07 (d, 1H, J=15.9 Hz, H-4), 3.58 (q, 2H,J=7.5 Hz, —OSO₂CH₂ CH₃), 3.45 (q, 2H, J=7.5 Hz, —OSO₂CH₂ CH₃), 3.35 (q,2H, J=7.5 Hz, —OSO₂ CH₂ CH₃), 1.67 (t, 3H, J=7.5 Hz, —OSO₂CH₂CH₃ ),1.63-1.56 (m, 6H, —OSO₂CH₂CH₃ ).

Compound 27: amorphous; ESI MS m/z: 559.1 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 7.71 (d, 1H, J=15.9 Hz, H-1), 7.65 (d, 1H, J=15.9 Hz, H-5),7.54-7.39 (m, 5H, aromatic ring H), 7.34-7.25 (m, 2H, aromatic ring H),7.09 (d, 1H, J=15.9 Hz, H-2), 7.06 (d, 1H, J=15.9 Hz, H-4), 3.40-3.01(m, 6H, —OSO₂CH₂ CH₃), 1.61-156 (m, 9H, —OSO₂CH₂CH₃ ).

Compound 40: oily syrups; C₂₁H₂₂O₆S, ESI-MS: m/z 403.2 [M+H]⁺; ¹H NMR(300 MHz, DMSO-d₆): 7.77-6.99 (11H, m, aromatic H and vinyl H), 3.80 (3Heach, s, OCH ₃), 3.77 (3H each, s, OCH ₃), 3.50 (2H, m, SO₂CH ₂CH₃),1.34 (3H, m, SO₂CH₂CH ₃).

Compound 43: pale yellow needles, mp 102-103° C.; C₂₃H₂₆O₁₀S₃, ESI-MS:m/z 559.1 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆): 7.89-7.35 (11H, m, aromaticH and vinyl H), 3.62 (4H, m, SO₂CH ₂CH₃×2), 3.53 (2H, q, J=7.5 Hz, SO₂CH₂CH₃), 1.37 (9H, m, SO₂CH₂CH ₃×3).

Compound 22: yellow solid; ESI MS m/z: 546.19 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 7.77 (s, 2H, benzylidene CH═), 7.50-7.28 (m, 8H, aromatic ringH), 3.92 (s, 4H, cyclohexyl-H-3,5), 3.35-3.25 (m, 6H, —OSO₂CH₂ CH₃),2.46 (d, 2H, J=6.6 Hz, —CH₂ ), 1.59-1.50 (m, 9H, —OSO₂CH₂CH₃ ), 0.86 (m,1H, cyclopropanyl CH), 0.50-0.44 (m, 2H, cyclopropanyl CH₂ ), 0.15-0.08(m, 2H, cyclopropanecarbonyl CH₂ ).

Compound 53: Yellow oily, ESI MS m/z: 481.0 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.68 (d, 2H, J=16.2 Hz, H-1,5), 7.63-7.54 (m, 2H, aromaticring H), 7.46-7.39 (m, 1H, aromatic ring H), 7.33-7.01 (m, 4H, aromaticring H; 2H, H-2,4 benzylidene CH═), 6.84-6.81 (m, 1H, aromatic ring H),2.95 (s, 12H, —OSO₂N(CH₃) ₂).

Compound 65: Amorphous, ESI MS m/z: 479.6 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ: 7.65 (d, 2H, J=16.0 Hz, H-1,5), 7.48 (br, 2H, aromatic ringH), 7.46-7.43 (m, 2H, aromatic ring H), 7.31 (d, 2H, J=8.4 Hz, aromaticring H), 6.99 (d, 2H, J=16.0 Hz, H-2,4), 3.36 (q, 4H, J=7.2 Hz, —OSO₂CH₂ CH₃), 2.37 (s, 6H, CH₃), 1.57 (t, 6H, J=7.2 Hz, —OSO₂CH₂ CH₃ ).

Compound 66: Orange-yellowish solid, ESI MS m/z: 401.6 [M+H]⁺; ¹H NMR(400 MHz, CDCl₃) δ: 7.67 (d, 1H, J=16.0 Hz, H-1), 7.63 (d, 1H, J=16.0Hz, H-5), 7.48-7.40 (m, 4H, aromatic ring H), 7.30 (d, 1H, J=8.4 Hz,aromatic ring H), 6.00 (d, 1H, J=16.0 Hz, H-2), 5.91 (d, 1H, J=16.0 Hz,H-4), 6.82 (d, 1H, J=8.4 Hz, aromatic ring H), 3.86 (s, 3H, OCH₃), 3.36(q, 2H, J=7.2 Hz, —OSO₂ CH₂CH₃ ), 2.37 (s, 3H, CH₃), 2.23 (s, 3H, CH₃),1.57 (t, 3H, J=7.2 Hz, —OSO₂CH₂ CH₃ ).

Compound 69: Brownish solid, ESI MS m/z: 479.6 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ: 7.94 (d, 2H, J=16.0 Hz, H-1,5), 7.65 (d, 2H, J=8.4 Hz,aromatic ring H), 7.14-7.12 (m, 4H, aromatic ring H), 6.92 (d, 2H,J=16.0 Hz, H-2,4), 3.28 (q, 4H, J=7.2 Hz, —OSO₂ CH₂ CH₃), 2.46 (s, 6H,—CH₃ ), 1.53 (t, 6H, J=7.2 Hz, —OSO₂CH₂ CH₃ ).

Compound 70: Light brownish solid, ESI MS m/z: 401.1 [M+H]⁺; ¹H NMR (400MHz, CDCl₃) δ: 7.99 (d, 1H, J=15.6 Hz, H-1), 7.91 (d, 1H, J=15.6 Hz,H-5), 7.65-7.61 (m, 2H, aromatic ring H), 7.14-7.12 (m, 3H, aromaticring H), 6.93 (d, 1H, J=15.6 Hz, H-2), 6.86 (d, 1H, J=15.6 Hz, H-4),6.77-6.74 (m, 1H, aromatic ring H), 3.82 (s, 3H, OCH₃), 3.29 (q, 2H,J=7.2 Hz, —OSO₂ CH₂ CH₃), 2.46 (s, 6H, CH₃), 1.53 (t, 3H, J=7.2 Hz,—OSO₂CH₂ CH₃ ).

Compound 74: Light yellowish solid, ESI MS m/z: 361.1 [M+H]⁺; ¹H NMR(400 MHz, CDCl₃) δ: 7.66 (d, 2H, J=16.0 Hz, H-1,5), 7.52-7.50 (m, 2H,aromatic ring H), 7.45-7.41 (m, 1H, aromatic ring H), 7.37-7.35 (m, 2H,aromatic ring H), 7.30-7.28 (m, 2H, aromatic ring H), 7.11-7.08 (m, 1H,aromatic ring H), 7.04 (d, 1H, J=16.0 Hz, H-2), 7.03 (d, 1H, J=16.0 Hz,H-4), 3.26 (q, 2H, J=7.6 Hz, —OSO₂ CH₂ CH₃), 1.55 (t, 3H, J=7.6 Hz,—OSO₂CH₂ CH₃ ).

Compound 77: Yellowish solid, ESI MS m/z: 427.1 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ: 7.68 (d, 2H, J=16.0 Hz, H-1,5), 7.53-7.51 (m, 3H, aromaticring H), 7.46-7.41 (m, 3H, aromatic ring H), 7.31-7.26 (m, 2H, aromaticring H), 7.05 (dd, 2H, J=16.0, 1.6 Hz, H-2,4), 3.31 (q, 2H, J=7.2 Hz,—OSO₂ CH₂ CH₃), 1.55 (t, 3H, J=7.2 Hz, —OSO₂CH₂ CH₃ ).

Compound 78 was synthesized in the manner similar to that in which1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one was synthesized. Aceticchloride (3 eq.) was used instead of ethanesulfonyl chloride. Afterstirring at room temperature for 3-4 h, the reaction mixture was pouredinto water, and extracted with CH₂Cl₂. The combined organic layers werewashed with water and then brine, dried over Na₂SO₄, filtered andconcentrated to give a light yellow solid crude product. Purificationwith CombiFlash chromatograph using n-hexane/EtOAc as eluent gave thedesired compound in quantitative yield. Light yellowish crystallinesolid, ESI MS m/z: 351.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ: 7.67 (d, 2H,J=16.0 Hz, H-1,5), 7.46-7.38 (m, 4H, aromatic ring H), 7.34-7.33 (m, 2H,aromatic ring H), 7.14-7.11 (m, 2H, aromatic ring H), 7.02 (d, 2H,J=16.0, H-2,4), 2.31 (s, 6H, —COCH₃ ).

Compounds 14 and 24 were synthesized in the manner similar to that inwhich 1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one was synthesized.Phosphorochloridic acid diethyl ester (˜10 eq. for Compound 14) orphosphorochloridic acid dimethyl ester (˜10 eq. for Compound 24) wereused instead of ethanesulfonyl chloride. After stirring at roomtemperature for 2 hour, the reaction mixture was poured into water, andextracted with CH₂Cl₂. The combined organic layers were washed withwater and then brine, dried over Na₂SO₄, filtered and concentrated togive a yellow solid crude product. Purification with CombiFlashchromatograph using CH₂Cl₂/MeOH as eluent gave the desired compounds inquantitative yield.

Compound 14: amorphous; ESI MS m/z: 539.22 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 7.69 (d, 2H, J=15.9 Hz, H-1,5), 7.51 (br. 2H, aromatic ringH), 7.46-7.39 (m, 4H, aromatic ring H), 7.32-7.28 (m, 2H, aromatic ringH), 7.09 (d, 2H, J=15.9 Hz, H-2,4), 4.32-4.22 (m, 8H, OCH₂ CH₃), 1.41(t, 12H, J=7.2 Hz, OCH₂CH₃ ).

Compound 24: amorphous, ESI MS m/z: 483.09 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 7.69 (d, 2H, J=15.9 Hz, H-1,5), 7.49-7.37 (m, 6H, aromaticring H), 7.28-7.25 (m, 2H, aromatic ring H), 7.06 (d, 2H, J=15.9 Hz,H-2,4), 3.91 (s, 6H, OCH₃ ), 3.87 (s, 6H, OCH₃ ).

Compounds 18, 23, 25, 32-34, and 37 were synthesized in the mannersimilar to that described above. Diethyl chlorophosphine (˜10-15 eq.)was used to prepare Compounds 18, 35, and 38, diphenyl chlorophosphine(˜10 eq.) was used to make Compound 23, and dimethyl chlorophosphine(˜10 eq.) was used to make Compounds 25, 32, and 33. After stirring atroom temperature for 2 h; the reaction mixture was poured into water andextracted with CH₂Cl₂. The organic layer was washed with water andbrine, dried over Na₂SO₄, filtered, and concentrated to give the crudecompound as a yellow solid. Purification with CombiFlash chromatographusing CH₂Cl₂/MeOH as the eluent gave the desired compound inquantitative yield.

Compound 18: amorphous; ESI MS m/z: 475.22 [M+H]⁺; ¹H NMR (300 MHz,CDCl₃) δ: 7.70 (d, 2H, J=15.9 Hz, H-1,5), 7.52 (d, 2H, J=0.9 Hz,aromatic ring H), 7.40-7.27 (m, 6H, aromatic ring H), 7.09 (d, 2H,J=15.9 Hz, H-2,4), 1.99-1.86 (m, 8H, CH₂ CH₃), 1.30-1.16 (m, 12H, CH₂CH₃).

Compound 23: yellow crystalline solid; ESI MS m/z: 667.61 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.95-7.88 (m, 8H, aromatic ring H), 7.76-7.67 (m,2H, aromatic ring H), 7.60 (d, 2H, J=15.9 Hz, H-1,5), 7.55-7.26 (m, 18H,aromatic ring H), 6.97 (d, 2H, J=15.9 Hz, H-2,4).

Compound 25: yellow crystalline solid; ESI MS m/z: 419.07 [M+H]⁺; ¹H NMR(300 MHz, CDCl₃) δ: 7.70 (d, 2H, J=15.9 Hz, H-1,5), 7.50 (d, 2H, J=1.5Hz, aromatic ring H), 7.45-7.37 (m, 4H, aromatic ring H), 7.30-7.26 (m,2H, aromatic ring H), 7.08 (d, 2H, J=15.9 Hz, H-2,4), 1.71 (s, 6H, CH₃),1.67 (s, 6H, CH₃).

Compound 32: light orange solid; ESI MS m/z: 381.14 [M+Na]⁺; ¹H NMR (400MHz, CD₃OD) δ: 8.09 (d, 1H, J=16.0 Hz, H-1), 7.65 (d, 1H, J=16.0 Hz,H-5), 7.26 (d, 1H, J=16.0 Hz, H-2), 7.15 (d, 1H, J=16.0 Hz, H-4), 7.23(t, 1H, J=8.0 Hz, aromatic ring H), 7.15-7.08 (m, 3H, aromatic ring H),6.85-6.81 (m, 2H, aromatic ring H), 6.70 (t, 1H, J=8.0 Hz, aromatic ringH), 1.26 (s, 6H, CH₃ ).

Compound 33: yellow solid; ESI MS m/z: 457.14 [M+Na]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 8.11 (d, 1H, J=16.0 Hz, H-1), 7.70 (d, 1H, J=16.0 Hz, H-5),7.56-7.54 (m, 1H, aromatic ring H), 7.44 (t, 1H, J=8.0 Hz, aromatic ringH), 7.28 (d, 1H, J=16.0 Hz, H-2), 7.15 (d, 1H, J=16.0 Hz, H-4),7.28-7.21 (m, 1H, aromatic ring H), 7.15-7.08 (m, 3H, aromatic ring H),6.84-6.82 (m, 1H, aromatic ring H), 6.70 (t, 1H, J=8.0 Hz, aromatic ringH), 1.72 (s, 3H, CH₃ ), 1.68 (s, 3H, CH₃ ), 1.27-1.19 (m, 6H, CH₃ ).

Compound 34: amorphous; ESI MS m/z: 489.08 [M−H]⁻; ¹H NMR (400 MHz,CDCl₃) δ: 8.14 (d, 1H, J=15.2 Hz, H-1), 7.73 (d, 1H, J=15.2 Hz, H-5),7.58-7.56 (m, 2H, aromatic ring H), 7.48-7.45 (m, 1H, aromatic ring H),7.33-7.28 (m, 3H, aromatic ring H), 7.17-7.14 (m, 1H, aromatic ring H),6.87-6.71 (m, 2H, H-2,4), 2.03-1.98 (m, 4H, CH₂ CH₃), 1.86-1.66 (m, 4H,CH₂ CH₃), 1.28-1.13 (m, 12H, CH₂CH₃ ).

Compound 37: amorphous; ESI MS m/z: 595.08 [M−H]⁻; ¹H NMR (400 MHz,CDCl₃) δ: 7.77 (d, 1H, J=15.6 Hz, H-5), 7.69 (d, 1H, J=15.6 Hz, H-1),7.56-7.53 (m, 2H, aromatic ring H), 7.44-7.40 (m, 2H, aromatic ring H),7.32-7.14 (m, 4H, aromatic ring H and H-2,4), 7.02-6.93 (m, 1H, aromaticring H), 2.02-1.89 (m, 12H, CH₂ CH₃), 1.26-1.06 (m, 18H, CH₂CH₃ ).

Compounds 28 and 29 were synthesized by the following procedure. Asolution of 1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one (synthesizedby the method illustrated in Scheme 4) (0.16 mmol) in acetonitrile wascooled to −78° C. To this was added Et₃N (20 eq.) and fresh distilledPOCl₃ (10 eq.) in acetonitriles. The resulting mixture was stirred at−78° C. for 2.5 hours or monitored by TLC. After completion, thereaction solution was warmed to 0° C. and water (˜1 mL) and pyridine(0.4 mL) were added. The resulting reaction mixture was stirred at 0° C.to room temperature for 1.5 hours and concentrated to dryness. The crudeas a mixture of Compounds 28 or 29 was purified by revered-phase columnchromatograph using C18 silica gel and MeOH/H₂O to afford the desiredproducts.

Compound 28: yellow solid; ESI MS m/z: 377.01 [M−H+MeOH]⁺; ¹H NMR (300MHz, D₂O) δ: 7.63 (d, 2H, J=15.9 Hz, H-1,5), 7.45-7.38 (m, 6H, aromaticH), 7.24-7.21 (m, 2H, aromatic H), 7.11 (d, 2H, J=15.9 Hz, H-2,4).

Compound 29: yellow solid; ESI MS m/z: 425.1 [M−H]⁺; ¹H NMR (300 MHz,D₂O) δ: 7.47 (d, 2H, J=15.9 Hz, H-1,5), 7.40-7.26 (m, 6H, aromatic H),7.21 (br, 2H, aromatic H), 6.95 (d, 2H, J=15.9 Hz, H-2,4).

Syntheses of Compounds 19, 35-36, 45-46, 48, 52, 54-59, and 76

Compounds 19, 35, 36, 67, 68, 71, and 72 were prepared from1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one or Compounds 20 and62-64 (Scheme 8). (2-Bromo-ethyl)-diethyl-amine hydrobromide (1 eq.) inDMF in the presence of K₂CO₃ (2.5 eq.) was used to prepare Compound 19.The reaction mixture was stirred at room temperature for 24 hours. Thesolid was filtered out and the filtrate was concentrated. The resultingresidue was diluted with CH₂Cl₂ and washed with water twice. The aqueouslayer was extracted with CH₂Cl₂ twice and dried over Na₂SO₄. Afterpurification by a CombiFlash chromatography system using Grace silicagel cartridge and CH₂Cl₂/MeOH, Compound 19 was obtained.

Compound 19: amorphous; ESI MS m/z: 366.30 [M+H]⁺; ¹H NMR (300 MHz,CD₃OD) δ: 7.74 (d, 1H, J=15.9 Hz, H-1), 7.73 (d, 1H, J=15.9 Hz, H-5),7.39-7.12 (m, 7H, aromatic H), 7.22 (d, 1H, J=15.9 Hz, H-2), 7.17 (d,1H, J=15.9 Hz, H-4), 7.63-7.02 and 6.89-6.84 (m, 1H, aromatic H), 4.25(t, 2H, J=5.4 Hz, OCH₂ CH₂N(CH₂CH₃)₂), 3.17 (t, 2H, J=5.4 Hz, OCH₂CH₂N(CH₂CH₃)₂), 2.93 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃)₂), 1.21 (t, 6H,J=7.2 Hz, OCH₂CH₂N(CH₂CH₃ )₂).

Compound 35: ESI MS m/z: 382.75 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 8.03(d, 1H, J=16.0 Hz, H-1), 7.34 (d, 1H, J=16.0 Hz, H-5), 7.17 (d, 1H,J=16.0 Hz, H-4), 7.10 (d, 1H, J=16.0 Hz, H-2), 7.23-7.07 (m, 4H,aromatic H), 6.98 (t, 1H, J=8.0 Hz, aromatic H), 6.87 (dd, 1H, J=1.6,8.4 Hz, aromatic H), 6.83 (dd, 1H, J=2.0, 8.4 Hz, aromatic H), 4.03 (t,2H, J=4.8, 9.6 Hz, OCH₂ CH₂N(CH₂CH₃)₂), 2.85 (t, 2H, J=4.8, 9.6 Hz,OCH₂CH₂ N(CH₂CH₃,)₂), 2.74 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃)₂), 1.13(t, 6H, J=7.2 Hz, OCH₂CH₂N(CH₂CH₃ )₂).

Compound 36: ESI MS m/z: 481.28 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 8.04(d, 1H, J=16.0 Hz, H-1), 7.74 (d, 1H, J=16.0 Hz, H-5), 7.25-7.15 (m, 6H,aromatic H, and H-4, 2), 7.02-6.95 (m, 2H, aromatic H), 6.89-6.85 (m,1H, aromatic H), 4.20 (dd, 2H, J=5.2, 4.8 Hz, OCH₂ CH₂N(CH₂CH₃)₂), 4.05(dd, 2H, J=5.2, 4.8 Hz, OCH₂ CH₂N(CH₂CH₃)₂), 3.10 (dd, 2H, J=4.8, 4.4Hz, OCH₂CH₂ N(CH₂CH₃,)₂), 2.92 (dd, 2H, J=4.8, 4.4 Hz, OCH₂CH₂N(CH₂CH₃)₂), 2.66 (m, 8H, OCH₂CH₂N(CH₂ CH₃)₂), 1.13 (m, 12H,OCH₂CH₂N(CH₂CH₃ )₂).

Compound 67: Amorphous, ESI MS m/z: 394.2 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.67 (d, 2H, J=15.6 Hz, H-1,5), 7.51-7.36 (m, 4H, aromaticring H), 7.08-6.70 (m, 4H, H-2,4 and aromatic ring H), 4.15 (t, 2H,J=5.2, Hz, OCH₂ CH₂N(CH₂CH₃)₂), 2.99 (t, 2H, J=5.2, Hz, OCH₂ CH₂N(CH₂CH₃)₂), 2.73-2.71 (m, 4H, OCH₂CH₂N(CH₂ CH₃)₂), 2.23 (s, 3H, CH₃),2.23-2.17 (m, 3H, CH₃), 1.13-1.09 (m, 6H, OCH₂CH₂N(CH₂ CH₃ )₂).

Compound 68: Amorphous, ESI MS m/z: 408.1 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.68 (d, 2H, J=15.2 Hz, H-1,5), 7.52-7.44 (m, 4H, aromaticring H), 7.07 (dd, 2H, J=15.2, 4.8 Hz, H-2,4), 6.96-6.92 (m, 2H,aromatic ring H), 4.18 (t, 2H, J=4.8, Hz, OCH₂ CH₂N(CH₂CH₃)₂), 3.85 (s,3H, OCH₃), 3.08 (t, 2H, J=5.2, Hz, OCH₂ CH₂ N(CH₂CH₃)₂), 2.80 (m, 4H,OCH₂CH₂N(CH₂ CH₃,)₂), 2.24 (s, 3H, CH₃), 2.20 (s, 3H, CH₃), 1.14 (t, 6H,J=6.8, Hz, OCH₂CH₂N(CH₂ CH₃ )₂).

Compound 71: light brownish solid, ESI MS m/z: 408.1 [M+H]⁺; ¹H NMR (400MHz, CD₃OD) δ: 8.03 (d, 2H, J=16.0 Hz, H-1,5), 7.74 (d, 2H, J=9.2 Hz,aromatic ring H), 7.07 (d, 2H, J=16.0, H-2,4), 6.81-6.75 (m, 3H,aromatic ring H), 6.64 (br, 1H, aromatic ring H), 4.11 (t, 2H, J=5.6,Hz, OCH₂ CH₂N(CH₂CH₃)₂), 3.80 (s, 3H, OCH₃), 2.90 (t, 2H, J=5.6, Hz,OCH₂ CH₂ N(CH₂CH₃)₂), 2.66 (m, 4H, OCH₂CH₂N(CH₂ CH₃,)₂), 2.45 (s, 6H,CH₃), 1.08 (t, 6H, J=7.2, Hz, OCH₂CH₂N(CH₂ CH₃ )₂).

Compound 72: yellowish solid, ESI MS m/z: 394.1 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 8.02 (dd, 2H, J=15.6, 4.8 Hz, H-1,5), 7.74-7.65 (m, 2H,aromatic ring H), 7.03 (d, 1H, J=15.6 Hz, H-2), 7.99 (d, 1H, J=15.6 Hz,H-4), 6.81 (br, 2H, aromatic ring H), 6.66-6.65 (m, 2H, aromatic ringH), 4.13 (t, 2H, J=5.6, Hz, OCH₂ CH₂N(CH₂CH₃)₂), 2.94 (t, 2H, J=5.5, Hz,OCH₂ CH₂ N(CH₂CH₃)₂), 2.73-2.66 (m, 4H, OCH₂CH₂N(CH₂ CH₃)₂), 2.45 (s,3H, CH₃), 2.40 (s, 3H, CH₃), 1.11-1.04 (m, 6H, OCH₂CH₂N(CH₂ CH₃ )₂).

Compounds 52, 54, and 58 were derived from1,5-bis-(3-hydroxy-phenyl)-penta-1,4-dien-3-one (synthesized by themethod shown in Scheme 4) by reacting with 1-bromobutane (3.0 eq. and1.2 eq. for compounds 52 and 54, respectively) or1-bromo-3-chloropropane (1.2 eq. for compound 58) in DMF in the presenceof potassium bicarbonate. The reaction mixture was stirred at roomtemperature (for compounds 52 and 58) or at 80° C. (for compound 58)overnight or with TLC monitoring. The solvent was evaporated underreduced pressure and the residue was partitioned in ethyl acetate andwater. The organic layer was washed with water twice and the aqueouswashings were extracted with ethyl acetate twice. After dried overNa₂SO₄, filtered, and concentrated, the oily crude was purified byCombiflash system using n-hexane/ethyl acetate eluent to give desiredcompounds 52, 54, and 58.

Compound 52: yellow crystalline solid; ESI MS m/z: 379.17 [M+H]⁺; ¹H NMR(400 MHz, CDCl₃) δ: 7.61 (d, 2H, J=16.0 Hz, H-1,5), 7.30-7.26 (m, 2H,aromatic ring H), 7.16-7.15 (m, 2H, aromatic ring H), 7.10 (br, 2H,aromatic ring H), 7.02 (dd, 2H, J=16.0, 2.0 Hz, H-2,4), 6.93-6.91 (m,2H, aromatic ring H), 3.97 (t, 4H, J=6.8 Hz, —OCH₂ CH₂CH₂CH₃), 1.79-1.72(m, 4H, OCH₂ CH₂ CH₂CH₃), 1.51-1.45 (m, 4H, OCH₂CH₂ CH₂ CH₃), 0.98-0.94(m, 6H, OCH₂CH₂CH₂ CH₃ ).

Compound 54: yellowish solid; ESI MS m/z: 323.08 [M+H]⁺; ¹H NMR (400MHz, CD₃OD) δ: 7.68 (dd, 2H, J=15.6, 4.4 Hz, H-1,5), 7.32-7.24 (m, 4H,aromatic ring H), 7.18-7.16 (m, 2H, aromatic ring H), 7.11 (br, 2H,aromatic ring H), 7.04 (dd, 2H, J=15.6, 4.0 Hz, H-2,4), 6.95-6.88 (m,2H, aromatic ring H), 3.99 (t, 2H, J=12.8, 6.8 Hz, OCH₂ CH₂CH₂CH₃),1.81-1.74 (m, 2H, OCH₂ CH₂ CH₂CH₃), 1.53-1.47 (m, 2H, OCH₂CH₂ CH₂ CH₃),0.99-0.96 (m, 3H, OCH₂CH₂CH₂ CH₃ ).

Compound 58: light yellow solid; ESI MS m/z: 329.12 [M+H]⁺; ¹H NMR (400MHz, CDCl₃) δ: 7.67 (d, 2H, J=16.0 Hz, H-1,5), 7.34-7.23 (m, 2H,aromatic H), 7.22 (d, 1H, J=8.0 Hz, aromatic ring H), 7.16 (d, 1H, J=8.0Hz, aromatic ring H), 7.13 (t, 1H, J=2.4 Hz, aromatic ring H), 7.11 (t,1H, J=2.0 Hz, aromatic ring H), 7.03 (dd, 2H, J=16.0, 2.4 Hz, H-2,4),6.97-6.94 (m, 1H, aromatic ring H), 6.91-6.88 (m, 1H, aromatic ring H),4.26 (t, 2H, J=6.0 Hz, —OCH₂ CH₂Cl), 3.82 (t, 2H, J=6.0 Hz, —OCH₂ CH₂Cl).

Compounds 55-57 and 59 were derived from the compound 58 (for compounds57 and 59) or the analogues of compound 58, in which the R₁ group wasreplaced with a 3-chloropropoxy (Scheme 8). To a solution of compound 58in DMF was added piperidine (˜4 eq). The resulting reaction mixture washeated to 80° C. and stirred overnight. After cooled to roomtemperature, the solvent was evaporated under reduced pressure and theresidue was partitioned in ethyl acetate and water. The organic phasewas washed with water twice and the aqueous washings were extracted withethyl acetate twice. After dried over Na₂SO₄, filtered, andconcentrated, the crude was purified by a Combiflash system usingmethylene chloride/methanol eluent, then crystallized from methanol togive compound 57. Amorphous, ESI MS m/z: 378.25 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.60 (d, 1H, J=16.0 Hz, H-1), 7.57 (d, 1H, J=16.0 Hz, H-5),7.27-7.22 (m, 2H, aromatic ring H), 7.15-7.11 (m, 2H, aromatic ring H),7.02 (t, 1H, J=2.0 Hz, aromatic ring H), 6.97 (d, 1H, J=16.0 Hz, H-2),6.92 (d, 1H, J=16.0 Hz, H-4), 6.88-6.84 (m, 3H, aromatic ring H), 4.13(t, 2H, J=5.6 Hz, OCH₂ CH₂N(CH₂CH₂)₂CH₂), 2.82 (t, 2H, J=5.6 Hz, OCH₂CH₂ N(CH₂CH₂)₂CH₂), 2.58 (br, 4H, OCH₂CH₂N(CH₂ CH₂,)₂CH₂), 1.68-1.63 (m,4H, OCH₂CH₂N(CH₂ CH₂ )₂CH₂), 1.48-1.45 (m, 2H, OCH₂CH₂N(CH₂CH₂,)₂ CH₂ ).

Compound 55 was synthesized in the same manner as described above byreaction of 3-chloropropoxy substituted1,5-bis-(3-substituted-phenyl)-penta-1,4-dien-3-one with diethylamine.Yield: 40%, amorphous, ESI MS m/z: 380.25 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.72 (d, 1H, J=16.0 Hz, H-1), 7.71 (d, 1H, J=16.0 Hz, H-5),7.35-7.25 (m, 4H, aromatic ring H), 7.23-7.21 (m, 1H, aromatic ring H),7.15 (d, 2H, J=16.0 Hz, H-2,4), 7.10 (t, 1H, J=2.0 Hz, aromatic ring H),7.01-6.98 (m, 1H, aromatic ring H), 6.86-6.83 (m, 1H, aromatic ring H),4.11 (t, 2H, J=6.4, Hz, OCH₂ CH₂CH₂N(CH₂CH₃)₂), 2.99-2.95 (m, 2H,OCH₂CH₂ CH₂ N(CH₂CH₃)₂), 2.88 (q, 4H, J=7.2 Hz, OCH₂CH₂CH₂N(CH₂ CH₃)₂),2.10-2.03 (m, 2H, OCH₂ CH₂ CH₂N(CH₂CH₃,)₂), 1.18 (t, 6H, J=7.2 Hz,OCH₂CH₂CH₂N(CH₂ CH₃ )₂).

Compound 56 was synthesized in the same manner as described above byreaction of 3-chloropropoxy substituted1,5-bis-(3-substituted-phenyl)-penta-1,4-dien-3-one with piperidine.Amorphous. ESI MS m/z: 392.25 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 7.72(d, 1H, J=16.0 Hz, H-1), 7.70 (d, 1H, J=16.0 Hz, H-5), 7.34-7.21 (m, 4H,aromatic ring H), 7.23 (d, 1H, J=16.0 Hz, H-2), 7.15 (d, 1H, J=16.0 Hz,H-4), 7.16-7.15 (m, 1H, aromatic ring H), 7.09 (t, 1H, J=2.4 Hz,aromatic ring H), 6.99-6.96 (m, 1H, aromatic ring H), 6.86-6.83 (m, 1H,aromatic ring H), 4.06 (t, 2H, J=6.4 Hz, —OCH₂ CH₂CH₂N(CH₂CH₂)₂CH₂),2.62-2.54 (m, 6H, —OCH₂CH₂ CH₂ N(CH₂ CH₂)₂CH₂), 2.05-1.98 (m, 2H, OCH₂CH₂ CH₂N(CH₂CH₂)₂CH₂), 1.66-1.61 (m, 4H, OCH₂CH₂CH₂N(CH₂ CH₂ )₂CH₂),1.51-1.49 (m, 2H, OCH₂CH₂CH₂N(CH₂ CH₂ ,)₂ CH₂ ).

Compound 59 was synthesized in the same manner as described above byreaction of compound 58 with piperidine. Yield: 35%, amorphous, ESI MSm/z: 380.25 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 7.72 (d, 1H, J=16.0 Hz,H-1), 7.70 (d, 1H, J=16.0 Hz, H-5), 7.35-7.21 (m, 5H, aromatic ring H),7.14 (d, 2H, J=16.0 Hz, H-2,4), 7.09 (t, 1H, J=2.0 Hz, aromatic ring H),7.02-6.99 (m, 1H, aromatic ring H), 6.86-6.83 (m, 1H, aromatic ring H),4.19 (t, 2H, J=5.6 Hz, OCH₂ CH₂N(CH₂CH₂)₂O), 3.71 (t, 4H, J=4.8 Hz,OCH₂CH₂N(CH₂ CH₂ )₂O), 2.84-2.81 (m, 2H, OCH₂ CH₂ N(CH₂CH₂)₂O), 2.61 (t,4H, J=4.8 Hz, OCH₂CH₂N(CH₂ CH₂,)₂O).

Compound 45 was synthesized by reacting Compound 16 with 1 eq. of(2-bromo-ethyl)-diethyl-amine hydrobromide in DMF in the presence ofK₂CO₃ (2.5 eq) (Scheme 7). The reaction mixture was stirred at roomtemperature overnight or with TLC monitoring. The solvent was evaporatedunder reduced pressure and the residue was partitioned in methylenechloride and water. The organic phase was washed with water three times(or to pH˜6-7) and the aqueous washings were extracted with methylenechloride twice. After dried over Na₂SO₄, filtered, and concentrated, theoily crude was purified by Combiflash system using methylenechloride/methanol eluent to give the desired product as a yellow solid.ESI MS m/z: 475.13 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 7.75 (d, 1H,J=11.6 Hz, benzylidene CH═), 7.38 (s, br, benzylidene CH═), 7.28 (s, br,1H, aromatic ring H), 7.11-6.84 (m, 7H, aromatic ring H), 5.04 (s, 2H,piperidin-4-one), 4.91 (s, 2H, piperidin-4-one), 4.21 (m, 2H, OCH₂CH₂N(CH₂CH₃)₂), 3.14 (m, 2H, OCH₂CH₂ N(CH₂CH₃,)₂), 2.87 (m, 4H,OCH₂CH₂N(CH₂ CH₃)₂), 1.17 (m, 6H, OCH₂CH₂N(CH₂CH₃ )₂), 0.87 (m, 1H,cyclopropanecarbonyl CH), 0.74 (m, 2H, cyclopropanecarbonyl CH₂ ), 0.62(m, 2H, cyclopropanecarbonyl CH₂ ).

Compound 46 was synthesized in the same manner as described above exceptthat 2 eq. (2-bromo-ethyl)-diethyl-amine hydrobromide and 5 eq. of K₂CO₃were used.

Compound 46: a yellow solid; ESI MS m/z: 574.21 [M+H]⁺; ¹H NMR (400 MHz,CD₃OD) δ: 7.75 (d, 1H, J=11.6 Hz, benzylidene CH═), 7.38 (s, br, 1H,benzylidene CH═), 7.28 (s, br, 1H, aromatic ring H), 7.11-6.84 (m, 7H,aromatic ring H), 5.05 (s, 2H, piperidin-4-one), 4.91 (s, 2H,piperidin-4-one), 4.49 (m, 2H, OCH₂ CH₂N(CH₂CH₃)₂, 3.86 (m, 2H, OCH₂CH₂N(CH₂CH₃)₂, 3.55 (q, 4H, J=7.6 Hz, OCH₂CH₂N(CH₂ CH₃)₂, 3.49 (t, 2H,J=6.4, Hz, OCH₂CH₂ N(CH₂CH₃,)₂, 2.87 (t, 2H, J=6.4 Hz, OCH₂CH₂N(CH₂CH₃,)₂, 2.62 (q, 4H, J=7.6 Hz, OCH₂CH₂N(CH₂ CH₃)₂, 1.38 (t, 6H,J=7.6 Hz, OCH₂CH₂N(CH₂CH₃ )₂, 1.05 (t, J=7.6 Hz, 6H, OCH₂CH₂N(CH₂CH₃ )₂,0.87 (m, 1H, cyclopropanecarbonyl CH), 0.74 (m, 2H, cyclopropanecarbonylCH₂ ), 0.64 (m, 2H, cyclopropanecarbonyl CH₂ ).

Compounds 48 and 76 were synthesized by reacting Compounds 19 and 59,respectively, with ethanesulfonyl chloride (˜1.3 eq.) and Et₃N (˜1.5eq.) in CH₂Cl₂ (Scheme 8). The reaction was carried out with stirring atroom temperature for 4-5 h. Upon the completion, the reaction mixturewas poured into water and washed with water twice. The aqueous layer wasextracted with CH₂Cl₂ twice. After purified with a Combiflash systemusing CH₂Cl₂/MeOH eluent, compound 48 was obtained in quantity as lightbrown solid. ESI MS m/z: 458.21 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 7.72(d, 1H, J=16.0 Hz, H-1), 7.71 (d, 1H, J=16.0 Hz, H-5), 7.64 (d, 1H,J=7.6 Hz, aromatic ring H), 7.61 (t, 1H, J=2.0 Hz, aromatic ring H),7.46 (t, 1H, J=8.0 Hz, aromatic ring H), 7.34-7.29 (m, 2H, aromatic ringH), 7.24-7.23 (m, 2H, aromatic ring H), 7.24 (d, 1H, J=16.0 Hz, H-2),7.19 (d, 1H, J=16.0 Hz, H-4), 6.99-6.97 (m, 1H, aromatic ring H), 4.11(t, 2H, J=5.6 Hz, OCH₂ CH₂N(CH₂CH₃)₂), 3.40 (q, 2H, J=7.6 Hz, SO₂CH₂CH₃), 2.91 (t, 2H, J=5.6 Hz, OCH₂CH₂ N(CH₂CH₃)₂), 2.67 (q, 4H, J=7.2 Hz,OCH₂CH₂N(CH₂ CH₃,)₂), 1.47 (t, 3H, J=7.6 Hz, SO₂CH₂CH₃ ), 1.08 (t, J=7.2Hz, 6H, OCH₂CH₂N(CH₂CH₃ )₂).

Compound 76: amorphous. ESI MS m/z: 470.1 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ: 7.67 (dd, 2H, J=16.0, 4.4 Hz, H-1,5), 7.54-7.50 (m, 2H,aromatic ring H), 7.43 (t, 1H, J=7.6 Hz, aromatic ring H), 7.32 (m, 2H,aromatic ring H), 7.20-7.15 (m, 2H, aromatic ring H), 7.05 (dd, 2H,J=16.0, 4.4 Hz, H-2,4), 6.95-6.92 (m, 1H, aromatic ring H), 4.30 (t, 2H,J=4.8 Hz, OCH₂ CH₂N(CH₂CH₂)₂CH₂), 3.30 (q, 2H, J=7.6 Hz, SO₂ CH₂ CH₃),3.01 (t, 2H, J=4.8 Hz, OCH₂ CH₂ N(CH₂CH₂)₂CH₂), 2.78 (br, 4H,OCH₂CH₂N(CH₂ CH₂,)₂CH₂), 1.76 (br, 4H, OCH₂CH₂N(CH₂ CH₂ )₂CH₂),1.56-1.52 (m, 5H, SO₂CH₂ CH₃ , and OCH2CH2N(CH₂CH₂)₂ CH₂ ).

Compound 47 was prepared by reacting compound 19 with succinic acid inacetone. See Scheme 9 above.

To a solution of succinic acid (0.397 g, 3.36 mmol) in acetone (9.5 mL)was added slowly with stirring compound 19 (1.23 g, 3.36 mmol) in 3-4 mLof acetone. During the addition, light yellow crystalline solid wasformed and precipitated out. After stirring at room temperature for 2 h,the reaction mixture was stored in a refrigerator overnight. The solidwas collected by filtration and washed with acetone to provide compound47 as a light yellow crystalline solid in 85% yield.

Compound 47: mp. 104-106° C.; ESI MS m/z: 366.30 [M-C₄H₆O₄+1]⁺; ¹H NMR(400 MHz, CD₃OD) δ: 7.72 (d, 1H, J=16.0 Hz, H-1), 7.70 (d, 1H, J=16.0Hz, H-5), 7.39-7.32 (m, 3H, aromatic H), 7.26 (d, 1H, J=16.0 Hz, H-2),7.23 (t, 1H, J=8.0 Hz, aromatic H), 7.14 (d, 1H, J=16.0 Hz, H-4),7.13-7.09 (m, 2H, aromatic H), 7.07-7.04 (m, 1H, aromatic H), 6.86-6.84(m, 1H, aromatic H), 4.37 (t, 2H, J=5.2 Hz, OCH₂ CH₂N(CH₂CH₃)₂, 3.50 (t,2H, J=5.2 Hz, OCH₂CH₂ N(CH₂CH₃)₂, 3.23 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH₂CH₃)₂, 2.49 (s, 4H, succinic acid —CH₂ CH₂ COOH), 1.32 (t, 6H, J=7.2 Hz,OCH₂CH₂N(CH₂CH₃ )₂.

Compound 49 was prepared from Compound 48 in a similar manner. SeeScheme 9 above.

Compound 49: brown viscous oil. ESI MS m/z: 458.21 [M-C₄H₆O₄+1]⁺; ¹H NMR(400 MHz, CD₃OD) δ: 7.77 (d, 2H, J=16.0 Hz, H-1,5), 7.67 (d, 1H, J=8.0Hz, aromatic ring H), 7.65 (t, 1H, J=2.0 Hz, aromatic ring H), 7.51 (t,1H, J=8.0 Hz, aromatic ring H), 7.38-7.33 (m, 4H, aromatic ring H), 7.30(d, 1H, J=16.0 Hz, H-2), 7.26 (d, 1H, J=16.0 Hz, H-4), 7.08-7.05 (m, 1H,aromatic ring H), 4.34 (t, 2H, J=5.2 Hz, OCH₂ CH₂N(CH₂CH₃)₂, 3.44 (t,2H, J=5.2 Hz, OCH₂CH₂ N(CH₂CH₃)₂, 3.42 (q, 2H, J=7.2 Hz, SO₂CH₂ CH₃),3.18 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH ₂CH₃,)₂, 2.49 (s, 4H, succinic acid—CH₂ CH₂ COOH), 1.49 (t, 3H, J=7.2 Hz, SO₂CH₂CH₃ ), 1.30 (t, 6H, J=7.2Hz, OCH₂CH₂N(CH₂CH₃ )₂.

Compound 50 was prepared by reacting Compound 48 with phosphoric acid(85% aqueous solution) in acetone. See Scheme 9 above.

To a solution of Compound 48 (0.28 g, 3.36 mmol) in acetone (0.5 mL) wasadded slowly with stirring a 85% H₃PO₄ aqueous solution (1 eq. of H₃PO₄,0.6 mmol) at 0° C. After stirring at room temperature for 1 h, thereaction mixture was stored in a refrigerator overnight. Afterevaporation of the solvent, the residue was dissolved in 7 mL of H₂O andthen frozen. Lyophilization gave Compound 50 as a yellow crystallinesolid at the yield of 95%.

Compound 50: ESI MS m/z: 458.21 [M−H₃PO₄+1]⁺; ¹H NMR (400 MHz, CD₃OD) δ:7.78 (d, 2H, J=16.0 Hz, H-1,5), 7.69 (d, 1H, J=7.6 Hz, aromatic ring H),7.65 (t, 1H, J=1.6 Hz, aromatic ring H), 7.51 (t, 1H, J=8.0 Hz, aromaticring H), 7.39-7.35 (m, 4H, aromatic ring H), 7.30 (d, 1H, J=16.0 Hz,H-2), 7.27 (d, 1H, J=16.0 Hz, H-4), 7.11-7.06 (m, 1H, aromatic ring H),4.43 (t, 2H, J=4.8 Hz, OCH₂ CH₂N(CH₂CH₃)₂, 3.60 (t, 2H, J=4.8 Hz,OCH₂CH₂ N(CH₂CH₃)₂, 3.42 (q, 2H, J=7.2 Hz, SO₂CH₂ CH₃), 3.33 (q, 4H,J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃,)₂, 1.49 (t, 3H, J=7.2 Hz, SO₂ CH₂ CH₃), 1.37(t, 6H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃)₂.

Compound 51 was prepared, as a bright yellow crystalline solid, fromCompound 19 in a similar manner. See Scheme 9 above.

Compound 51: ESI MS m/z: 366.30 [M−H₃PO₄+1]⁺; ¹H NMR (400 MHz, CD₃OD) δ:7.74 (d, 1H, J=16.0 Hz, H-1), 7.72 (d, 1H, J=16.0 Hz, H-5), 7.39-7.35(m, 3H, aromatic H), 7.28 (d, 1H, J=16.0 Hz, H-2), 7.24 (t, 1H, J=7.6Hz, aromatic H), 7.16 (d, 1H, J=16.0 Hz, H-4), 7.17-7.15 (m, 1H,aromatic H), 7.11-7.06 (m, 2H, aromatic H), 6.87-6.84 (m, 1H, aromaticH), 4.42 (t, 2H, J=4.8 Hz, OCH₂ CH₂N(CH₂CH₃)₂, 3.61 (t, 2H, J=4.8 Hz,OCH₂CH₂ N(CH₂CH₃)₂, 3.33 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃)₂, 1.37 (t,6H, J=7.2 Hz, OCH₂CH₂N(CH₂CH₃ )₂.

Compound 79 was prepared as a yellow crystalline solid by reactingCompound 48 with 2.0 M hydrochloride ethyl ether solution.

To a solution of Compound 48 in methanol was added slowly with stirringa 2.0 M hydrochloride ethyl ether solution at 0° C. After stirring atroom temperature for 1 h, the reaction mixture was stored in arefrigerator overnight. The solvent was evaporated and the residue waswashed with t-buty methyl ether three times. Lyophilization gaveCompound 79 as a yellow crystalline solid in quantitative yield.

To a solution of Compound 48 in methanol was added slowly with stirringa 2.0 M hydrochloride ethyl ether solution at 0° C. After stirring atroom temperature for 1 h, the reaction mixture was stored in arefrigerator overnight. The solvent was evaporated and the residue waswashed with t-buty methyl ether three times. Lyophilization gaveCompound 79 as a yellow crystalline solid in quantitative yield. ESI MSm/z: 458.21 [M−HCl+1]⁺; ¹H NMR (400 MHz, CD₃OD) δ: 7.80 (d, 2H, J=16.0Hz, H-1,5), 7.71 (d, 1H, J=8.0 Hz, aromatic ring H), 7.67 (t, 1H, J=2.4Hz, aromatic ring H), 7.53 (t, 1H, J=8.0 Hz, aromatic ring H), 7.42-7.41(m, 2H, aromatic ring H), 7.39-7.37 (m, 2H, aromatic ring H), 7.32 (d,1H, J=16.0 Hz, H-2), 7.29 (d, 1H, J=16.0 Hz, H-4), 7.13-7.10 (m, 1H,aromatic ring H), 4.44 (t, 2H, J=4.8 Hz, OCH₂ CH₂N(CH₂CH₃)₂, 3.65 (t,2H, J=4.8 Hz, OCH₂ CH₂ N(CH₂CH₃)₂, 3.43 (q, 2H, J=7.6 Hz, SO₂ CH₂ CH₃),3.37 (q, 4H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃,)₂, 1.51 (t, 3H, J=7.6 Hz,SO₂CH₂ CH₃ ), 1.40 (t, 6H, J=7.2 Hz, OCH₂CH₂N(CH₂ CH₃ )₂.

Syntheses of Compounds 60 and 61

Compounds 60 and 61 were synthesized from α-bromomethyl-tolunitril. To asolution of α-bromomethyl-tolunitril (15.3 mmol) in toluene (30 mL) wasadded DIBAL-H in THF (1.0 M, 1.4 eq.) at 0° C. within 30 minutes. Afterstirring at 0° C. for 2 h, the reaction mixture was poured into amixture of 40 mL of methylene chloride and 100 mL of 10% HCl. Theresulting mixture was stirred for 1 h and the organic layer was washedwith water and then brine, and the aqueous was extracted with methylenechloride twice. After dried over Na₂SO₄, filtered, and concentrated, theobtained semi-oily product was stored in a refrigerator to give3-(bromomethyl)benzaldehyde as a white crystalline solid in quantitativeyield. Reaction of the resulting compound with acetone in ethanolfollowing the procedure descripted in Scheme 4 yielded Compound 61 as alight yellow crystalline solid. ESI MS m/z: 420.9 [M+H]⁺; ¹H NMR (400MHz, CDCl₃) δ: 7.71 (d, 2H, J=16.0 Hz, H-1,5), 7.63-7.60 (m, 2H,aromatic ring H), 7.54-7.50 (m, 2H, aromatic ring H), 7.44-7.37 (m, 4H,aromatic ring H), 7.08 (d, 2H, J=16.0 Hz, H-2,4), 4.50 (s, 4H, —CH₂ Br).

Compound 60 was obtained by reacting Compound 61 in CH₂Cl₂ with amixture of ethanesulfonyl chloride (2 eq.), sodium sulfite (4 eq.),sodium bicarbonate (4 eq.) in water. The resulting mixture was stirredat 35-36° C. overnight. The reaction mixture was diluted with methylenechloride, washed with water then brine, and dried over Na₂SO₄. The crudeproduct was purified with a Combiflash system using n-hexanes/EtOAceluent to afford the desired product as light yellow solid. Yield: 44%.ESI MS m/z: 447.1[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ: 7.55 (d, 2H, J=16.0Hz, H-1,5), 7.53 (br, 2H, aromatic H), 7.47-7.25 (m, 2H, aromatic ringH), 7.42-7.38 (m, 4H, aromatic ring H), 6.70 (d, 2H, J=16.0 Hz, H-2,4),4.57 (s, 4H, —CH₂ SO₂CH₂CH₃), 2.82-2.70 (m, 4H, —CH₂SO₂ CH₂ CH₃), 1.30(t, 6H, J=7.2 Hz, —CH₂SO₂ CH₂CH₃ ),

Biological Assays

Inhibitory Effects on Proliferation of Human Lernert's T Cell LymphomaKT-3

Inhibitory effects of compounds of this invention on the growth of humanLennert's T cell lymphoma KT-3 cells, an IL-6-dependent cell line, weredetermined as described below. Briefly, KT-3 cells were transferred intowells of 96-well plates (2.5×10³/well) and cultured in RPMI-1640 medium(GIBCO) containing penicillin (25 U/ml), streptomycin (25 μg/ml), 10%heat-inactivated fetal bovine serum (FBS), and hIL-6 (2.5 ng/ml, R&Dsystems). To experimental wells, test compounds at variousconcentrations (in triplicates) were added immediately after the cellswere plated. To the control wells, an equal volume of vehicle, DMSO(0.1% v/v), was added. After incubating the cells with the compounds orin vehicle for 48 hours, cell viability was assessed using a CellTiterGlo Luminescent Cell Viability Assay Kit (Promega, Madison, Wis.). Theresultant luminescence was quantified using Microplate Luminometer LB96V(EG&G BERTH HOLD) according to the manufacturer's protocol. The cellsurviving rate was calculated by dividing the luminescence value ofcompound-treated cells with that of vehicle-treated cells.

All of Compounds 1-79 exhibited inhibitory effect on IL-6-induced KT-3cell proliferation. Some compounds, i.e., Compounds 6, 16, 18, 19, 26,27, 35, 36, 45-51, 56, 57 and 79 unexpectedly had IC₅₀ values (theconcentration at which a compound suppresses cell growth by 50%) equalto or even lower than 0.05 μM. In addition, all compounds 1-79 inhibitedKT-3 cell proliferation in a dose-dependent manner.

Inhibition of Growth of Various Human Tumor Cell Lines

The inhibitory effects of Compound 6 were determined as described belowon the growth of colon cancer cells (HCT116, HT29, SW480, and SW620),colon adenocarcinoma cells (Colo205), prostate adenocarcinoma cells(PC-3 and Dul145), prostate carcinoma cells (CWR22RV and LNCap),non-small cell lung carcinoma cells (NCI-H1299), lung adnocarcinomacells (A549), large cell lung carcinoma cells (NCI-H460), breastmetastatic carcinoma cells (MDA-MB-453), breast ductal carcinoma cells(T-47D), breast adnocarcinoma cells (MCF7), hepatocellular carcinomacells (Huh-7 and HepG2), pancreatic carcinoma cells (PANC-1), cervixadnocarcinoma cells (Hela), IL-6-dependent Lennert's T-cell lymphomacells (KT-3), IL-6-dependent multiple myeloma cells (INA-6), multiplemyeloma cells (KMM-1 and U266), myeloid leukemia cells (HL-60), andT-cell leukemia cells (Jurkat). Briefly, tumor cells were seeded atdensities ranging from 1×10³ to 4×10³/well in 96-well Microtest IIItissue culture plates (Falcon, N.J.). After cultured in DMEM (GIBCO)containing penicillin (25 units/milliliter), streptomycin (25micrograms/milliliter), and 10% heat-inactivated FBS for 12 hours, cellswere treated with various concentrations ranging from 0 to 5 μM ofcompound 6 for 72 h. The viability of the tumor cells was then assessedusing the tetrazolium-based calorimetric assay (MTT) as described in Suet al., J Mol Cell Cardiol. 1998; 30:587-598. The cells were incubatedwith the MTT dye (5 mg/mL) at 37° C. for 3 hours. The resultant formazancrystals were solubilized with MTT lysis buffer (50% DMF, 24 mM HCl, 2%Acetic Acid, 5% SDS) and the absorbance at 595 nm was measured using aBenchmark microplate reader (BIO-RAD, Hercules, Calif.). The cellssurviving rate was calculated by the absorbance value ofcompound-treated cells vs. that of vehicle-treated cells.

Compound 6 exhibited dose-dependent inhibitory effects on the growth ofall tumor cells tested and with IC₅₀ values ranging from 0.02 to 5.5 μM.

Western Blot Analysis of MRG, STAT1, STAT3, and STAT5 Proteins in KT-3Cells

Western Blot analysis was performed as described in Kawashima et al., J.Immunol. 2001, 167:3652-3660, with minor modifications. Briefly, KT-3cells were treated with 1, 5, or 20 μM of compound 6 or vehicle alone inthe above described IL-6-containing medium. At various time points afterincubation (i.e., 0.5, 1, 3, and 6 hours after the treatment), cellswere harvested and lysed using a lysis buffer (1.0% Triton X-100, 50 mMTris-HCl (pH 7.5), 0.1 mM EDTA, 150 mM NaCl, 200 μM Na₃VO₄, 50 mM NaF, 1mM dithiothreitol, 0.4 mM phenylmethylsulfonyl fluoride, 3 μg/ml ofaprotinin, 2 μg/ml of pepstatin A, 1 μg/ml of leupeptin) at 2×10⁷cells/ml on ice for 30 min. Cell lysates were harvested bycentrifugation at 12,000×g for 15 min. The samples (1×10⁵ cellequivalent/lane) were subjected to sodium dodecyl sulfate-polyacrylamidegel electrophoresis and subsequently transferred onto Immobilon filters(Millipore). After blocked with 5% BSA, the filter was probed withanti-pSTAT3, STAT3, STAT5, STAT1, MRG or Actin Abs. The rabbitpolyclonal anti-STAT3, anti-STAT5, anti-STAT1, and anti-Actin antibodies(Abs), and mouse monoclonal anti-pSTAT3 Ab (B-7) were obtained fromSanta Cruz Biotechnology. Affinity purified anti-MRG Ab was prepared asdescribed previously in Hirose et al., J. Biol. Chem. 276:5821-5828. Thefilter was further incubated with HRP-conjugated secondary antibodies.Finally, the proteins were visualized using the EnhancedChemiluminescence (ECL) system (Amersham).

The results show that Compound 6 down-regulated MRG (MgcRacGAP, anevolutionarily conserved GTPase-activating) protein, STAT3, and STAT5proteins and inhibited phosphorylation of STAT3 protein in KT-3 cells ina time and dose-dependent manner. In contrast, this compound had noeffect on the expression of STAT1 in the KT3 cells under the testingcondition.

Inhibitory Effect on Human Colon Cancer (HCT-116) in a Mouse XenograftModel

The in vivo experiments were carried out under an Institutional AnimalCare and Use Committee-approved protocol and followed Institutionalguidelines for proper and humane use of animals in research.Six-week-old female athymic nude mice were purchased from the HarlanLaboratory. Each of eight mice was injected subcutaneously with 1×10⁶HCT-116 cells mixed with Matrigel (BD Bioscience) at the left flank.After injection, tumors were allowed to grow for 5 days into a palpablesize (volume ˜100 mm³) before treatment. Mice were randomized into twogroups (n=4 per group). The control group was injected with a vehiclesolution only (i.e., 10% DMSO and 90% corn oil), while the experimentalgroup received a daily injection (i.p.) of compound 6 (at a dose of 40mg/kg body weight, dissolved in the vehicle solution) from days 0 (theinitiation day of injection) to 4 and from days 7 to 10. The sizes oftumors were measured twice a week using a Vernier caliper and calculatedusing the following formula: length×width×height×0.5236, as described inRockwell et al., J Natl. Cancer Inst. 1972; 49:735-747.

The result shows that Compound 6 inhibited human colon cancer growth inthe xenograft mouse model.

OTHER EMBODIMENTS

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the scope of thefollowing claims.

The invention claimed is:
 1. A method for treating a signal transduction and transcription (STAT) protein-associated condition, the method comprising administering to a subject in need thereof an effective amount of a compound of formula (I):

wherein each of X and Y, independently, is H, alkyl, or halo; and each of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′), independently, is H, alkyl, halo, OH, R_(c)—O—, R_(d)S(O)₂—O—, (R_(d))₂P(O)—O—, or (R_(d)O)₂P(O)—O—, R_(c) being unsubstituted alkyl or alkyl substituted with halo, OH, alkoxy, amino, or cycloalkyl, and R_(d) being H, OH, C₂-C₁₀ alkyl, or alkoxy; in which at least one of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′) is R_(d)S(O)₂—O—, (R_(d))₂P(O)—O—, or (R_(d)O)₂P(O)—O—, and the STAT protein-associated condition is lymphoma lung cancer, breast cancer, pancreatic cancer, cervical cancer, leukemia, or multiple myeloma.
 2. The method of claim 1, wherein the STAT protein-associated condition is pancreatic cancer, lung cancer, breast cancer, lymphoma, or leukemia.
 3. The method of claim 1, wherein the compound is one of the following compounds:


4. The method of claim 3, wherein the STAT protein-associated condition is pancreatic cancer, lung cancer, breast cancer, lymphoma, or leukemia.
 5. A method for treating a STAT protein-associated condition, the method comprising administering to a subject in need thereof an effective amount of one of the following compounds:

wherein the STAT protein-associated condition is lymphoma, lung cancer, breast cancer, pancreatic cancer, cervical cancer, leukemia, or multiple myeloma.
 6. The method of claim 5, wherein the compound is one of the following compounds:


7. The method of claim 5, wherein the STAT protein-associated condition is pancreatic cancer, lung cancer, breast cancer, lymphoma, or leukemia.
 8. The method of claim 7, wherein the compound is one of the following compounds:


9. The method of claim 5, wherein the STAT protein-associated condition is pancreatic cancer.
 10. The method of claim 9, wherein the compound is one of the following compounds:


11. The method of claim 5, wherein the STAT protein-associated condition is lung cancer.
 12. The method of claim 11, wherein the compound is one of the following compounds:


13. The method of claim 5, wherein the STAT protein-associated condition is breast cancer.
 14. The method of claim 13, wherein the compound is one of the following compounds:


15. The method of claim 5, wherein the STAT protein-associated condition is cervical cancer.
 16. The method of claim 15, wherein the compound is one of the following compounds:


17. The method of claim 5, wherein the STAT protein-associated condition is multiple myeloma, lymphoma, or leukemia.
 18. The method of claim 17, wherein the compound is one of the following compounds:


19. A method for deactivating a STAT protein, the method comprising contacting the STAT protein with an effective amount of a compound of formula (I):

wherein each of X and Y, independently, is H, alkyl, or halo; and each of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′), independently, is H, alkyl, halo, OH, R_(c)—O—, R_(d)S(O)₂—O—, (R_(d))₂P(O)—O—, or (R_(d)O)₂P(O)—O—, R_(c) being unsubstituted alkyl or alkyl substituted with halo, OH, alkoxy, amino, or cycloalkyl, and R_(d) being H, OH, C₂-C₁₀ alkyl, or alkoxy; in which at least one of R¹, R², R³, R⁴, R⁵, R^(1′), R^(2′), R^(3′), R^(4′), and R^(5′) is R_(d)S(O)₂—O—, (R_(d))₂P(O)—O—, or (R_(d)O)₂P(O)—O—. 